CN114870772B - Decellularization reaction kettle device and decellularization method - Google Patents
Decellularization reaction kettle device and decellularization method Download PDFInfo
- Publication number
- CN114870772B CN114870772B CN202210231426.2A CN202210231426A CN114870772B CN 114870772 B CN114870772 B CN 114870772B CN 202210231426 A CN202210231426 A CN 202210231426A CN 114870772 B CN114870772 B CN 114870772B
- Authority
- CN
- China
- Prior art keywords
- reagent
- box
- decellularized
- inner box
- controller
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
- A61L27/3629—Intestinal tissue, e.g. small intestinal submucosa
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J19/18—Stationary reactors having moving elements inside
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Dermatology (AREA)
- Medicinal Chemistry (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Urology & Nephrology (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a cell-removing reaction kettle device which comprises an outer box, an inner box, a material layering mechanism, a rotary driving mechanism, a temperature control mechanism, a drying mechanism, a sterilization mechanism, a waste liquid removing mechanism and a controller, wherein the outer box is connected with the inner box through a pipeline; the inner box is arranged in the outer box, the material layering mechanism is arranged in the inner box, and the top of the inner box is of an open structure; the inner box is communicated with a decellularized reagent liquid storage box for providing a decellularized reagent and a cleaning reagent liquid storage box for providing a cleaning reagent, and the decellularized reagent and the cleaning reagent can enter the material layering mechanism through the inner box and contact with the membrane biological materials in the material layering mechanism; the rotary driving mechanism is connected with the material layering mechanism and can drive the material layering mechanism to rotate; the rotary driving mechanism, the drying mechanism, the sterilizing mechanism and the waste liquid removing mechanism are all electrically connected with the controller. The device reduces the labor cost, saves the time of cell removal and improves the working efficiency.
Description
Technical Field
The invention relates to the technical field of decellularization, in particular to a decellularization reaction kettle device and a decellularization method.
Background
In recent years, more and more decellularized tissue engineering scaffold materials are applied to soft tissue defect repair, and are used for performing decellularization treatment on dermis tissues, small intestine submucosa or vascular tissues of human or animals by a physical, chemical or enzymatic method to obtain extracellular matrixes, so that a scaffold with a porous three-dimensional structure is formed, and a good microenvironment is provided for adhesion, growth and proliferation of host cells. The extracellular matrix obtained by decellularization treatment is mainly collagen, has high affinity to host tissue cells, excellent biomechanical property, can induce tissue regeneration, is used for repairing the defect of soft tissues, and can effectively recover the appearance and the function of the defect tissues.
At present, the conventional decellularization treatment is mainly finished in a reaction container through oscillation, and the process of the decellularization treatment needs to be controlled manually, so that the time consumption of the decellularization treatment process is long, the procedure is complicated, and the labor cost is high.
Therefore, how to provide a cell-removing reaction kettle device, which can save time of cell-removing treatment, improve efficiency and effectively reduce labor cost is a technical problem that needs to be solved by those skilled in the art.
Disclosure of Invention
Accordingly, the present invention is directed to a decellularized reactor device, which can save time for decellularizing treatment, improve efficiency, and effectively reduce labor cost.
It is also an object of the present invention to provide a method of decellularizing.
In order to achieve the above purpose, the present invention provides the following technical solutions:
The cell-removing reaction kettle device comprises an outer box, an inner box, a material layering mechanism, a rotary driving mechanism, a temperature control mechanism, a drying mechanism, a sterilization mechanism, a waste liquid removing mechanism and a controller;
the inner box is arranged in the outer box, the material layering mechanism is arranged in the inner box, and the top of the inner box is of an open structure;
The inner box is communicated with a decellularized reagent liquid storage box for providing a decellularized reagent and a cleaning reagent liquid storage box for providing a cleaning reagent, and the decellularized reagent and the cleaning reagent can enter the material layering mechanism through the inner box and contact with membrane biological materials in the material layering mechanism;
The rotary driving mechanism is connected with the material layering mechanism and can drive the material layering mechanism to rotate;
The rotary driving mechanism, the drying mechanism, the sterilization mechanism and the waste liquid removing mechanism are all electrically connected with the controller.
Preferably, the material layering mechanism comprises at least two layers of storage frames for storing the membrane biological materials, and any two adjacent layers of storage frames are detachably connected;
a plurality of through holes are formed in any one of the storage frames, and the through holes are used for communicating the inner cavity of the inner box with the inner cavity of the storage frame;
the upper cover of the storage frame is further arranged on the storage frame arranged on the top layer of the material layering mechanism.
Preferably, at least one of the decellularized reagent liquid storage tank and the cleaning reagent liquid storage tank is arranged;
Any one of the decellularized reagent liquid storage tanks is communicated with the inner cavity of the inner tank through a first liquid inlet pipe;
any one of the cleaning reagent liquid storage tanks is communicated with the inner cavity of the inner tank through a second liquid inlet pipe.
Preferably, the device further comprises a liquid inlet which is communicated with the inner cavity of the inner box and the first liquid inlet pipe, and the inner cavity of the inner box and the second liquid inlet pipe, wherein the first liquid inlet pipe and the second liquid inlet pipe are communicated with the liquid inlet and are communicated with the inner cavity of the inner box through the liquid inlet.
Preferably, the waste liquid removing mechanism comprises a liquid discharge pipe communicated with the inner cavity of the inner box and a liquid discharge valve arranged in the liquid discharge pipe, and the liquid discharge valve is electrically connected with the controller.
Preferably, the temperature control mechanism comprises a cooling pipeline arranged on the inner box, and a water inlet and a water outlet which are arranged on the inner box and are respectively communicated with two ends of the cooling pipeline;
A water inlet valve is arranged at the water inlet, a water outlet valve is arranged at the water outlet, and the water inlet valve and the water outlet valve are electrically connected with the controller;
The water inlet is arranged at the lower part of the inner box, and the water outlet is arranged at the upper part of the inner box;
Preferably, the side plate of the inner box comprises an inner side plate and an outer side plate, an accommodating space is arranged between the inner side plate and the outer side plate, and the cooling pipeline is arranged in the accommodating space.
Preferably, the accommodating space is further filled with a heat insulation material, the heat insulation material is arranged close to the inner wall of the outer side plate, and the cooling pipeline is arranged close to the inner wall of the inner side plate.
Preferably, the drying mechanism is a circulating fan, one end of the circulating fan is arranged on the outer box, and the other end of the circulating fan is arranged inside the outer box.
Preferably, the sterilization mechanism is an ultraviolet sterilization lamp.
Preferably, the device further comprises a liquid level sensor arranged in the inner box, and the liquid level sensor is electrically connected with the controller.
Preferably, the device further comprises a supporting mechanism for supporting the outer box;
The supporting mechanism comprises a hollow supporting box body and rollers arranged at the bottom of the supporting box body, and the rotary driving mechanism is arranged in the supporting box body.
A decellularization method comprising:
S100: placing a membrane biological material in a material layering mechanism, and controlling a decellularized reagent liquid storage tank to provide a decellularized reagent for an inner tank, wherein the decellularized reagent enters the material layering mechanism to be contacted with the membrane biological material;
S200: controlling the rotation driving mechanism to rotate so as to drive the material layering mechanism to rotate, and simultaneously controlling the temperature of the inner box to be a preset temperature;
s300: when the reaction of the membrane biological material and the decellularized reagent reaches a first preset time, controlling a waste liquid removing mechanism to discharge the first waste liquid of the inner box;
S400: controlling a cleaning reagent liquid storage tank to supply cleaning reagent to the inner tank;
s500: controlling the rotary driving mechanism to be started, cleaning the membrane biological materials, and controlling the waste liquid cleaning mechanism to discharge second waste liquid of the inner box after the cleaning reaches a second preset time;
S600: and taking the membrane biological material out of the material layering mechanism, controlling the drying mechanism to dry the inner box, and controlling the sterilizing mechanism to sterilize the dried inner box.
Preferably, the rotating speed range of the rotary driving mechanism is 200-300rpm/min, the first preset time is 1-2h, and the temperature of the inner box is 25-40 ℃.
Compared with the prior art, the technical scheme shows that the cell-removing reaction kettle device carries out cell-removing treatment through an automatic program in the cell-removing treatment process of the membrane biological material, so that the cost of manual liquid adding, liquid changing, cleaning, liquid discharging, drying, sterilization and the like is greatly reduced, the cell-removing time is saved, the working efficiency is improved, and favorable conditions are created for the large-scale production and application of the membrane biological material.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the description of the embodiments or the prior art are briefly introduced below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic structural diagram of a cell-free reactor device according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a material layering mechanism according to an embodiment of the present invention;
FIG. 3 is a schematic view of a storage frame of a material layering mechanism according to an embodiment of the present invention;
FIG. 4 is a schematic flow chart of a method for decellularizing according to an embodiment of the invention.
Wherein, each part name is as follows:
100 is an outer box, 101 is an outer box upper cover, 102 is an outer box handle, 103 is a drying mechanism, 104 is a sterilizing mechanism, 105 is an air inlet, 106 is an air outlet, and 107 is a liquid inlet
200 Is an inner box, 201 is an inner side plate, 202 is an outer side plate, 203 is a cooling pipeline, 204 is a water inlet, 205 is a water outlet, 206 is a heat insulation material, 207 is an electric heating pipe, 208 is an air duct, 209 is a liquid discharge pipe, 2091 is a liquid discharge valve, 210 is a liquid level sensor,
300 Is a material layering mechanism, 301 is a storage frame, 3011 is a through hole, 302 is a storage frame upper cover, 303 is a storage frame handle, 304 is a buckle,
400 Is a rotary driving mechanism, 500 is a supporting mechanism, 501 is a roller, 502 is a bearing, 600 is a decellularized reagent liquid storage tank, 601 is a first decellularized reagent liquid storage tank, 602 is a second decellularized reagent liquid storage tank, 603 is a third decellularized reagent liquid storage tank, 604 is a fourth decellularized reagent liquid storage tank, 605 is a fifth decellularized reagent liquid storage tank, 606 is a first liquid inlet pipe, 607 is a first vacuum pump, 700 is a cleaning reagent liquid storage tank, 701 is a second vacuum pump, and 800 is a controller.
Detailed Description
Accordingly, the core of the present invention is to provide a decellularized reactor device, which can save time of decellularized treatment, improve efficiency and effectively reduce labor cost.
Another core of the present invention is also to provide a decellularization method.
For a better understanding of the present invention, reference is made to fig. 1 to 4, for a further detailed description of the present invention, taken in conjunction with the accompanying drawings and detailed description.
Referring to fig. 1, the cell-removing reactor device disclosed in the embodiment of the present invention includes an outer case 100, an inner case 200, a material layering mechanism 300, a rotation driving mechanism 400, a temperature control mechanism, a drying mechanism 103, a sterilization mechanism 104, a waste liquid removing mechanism and a controller 800, wherein the inner case 200 is disposed inside the outer case 100, the material layering mechanism 300 is disposed inside the inner case 200, and the top of the inner case 200 is of an open structure; the inner box 200 is communicated with a decellularization reagent liquid storage box 600 for providing a decellularization reagent and a cleaning reagent liquid storage box 700 for providing a cleaning reagent, and the decellularization reagent and the cleaning reagent can enter the material layering mechanism 300 through the inner box 200 and contact with membrane biological materials in the material layering mechanism 300; the rotary driving mechanism 400 is connected with the material layering mechanism 300, and the rotary driving mechanism 400 can drive the material layering mechanism 300 to rotate; the rotary drive mechanism 400, the drying mechanism 103, the sterilization mechanism 104, and the waste liquid removal mechanism are all electrically connected to the controller 800.
When the cell removal treatment is performed on the membrane biological material, the membrane biological material is firstly placed in the material layering mechanism 300 in the inner box 200, then the controller 800 is started, the cell removal reagent is controlled by the controller 800 to be supplied to the inner box 200 from the cell removal reagent liquid storage box 600, the rotation driving mechanism 400 is controlled by the controller 800 to rotate so as to drive the material layering mechanism 300 to rotate, the membrane biological material is fully contacted with the cell removal reagent in the process of rotating along with the material layering mechanism 300, and meanwhile, the temperature of the temperature control mechanism is controlled by the controller 800, so that the temperature of the inner box 200 is kept within a preset temperature range;
after the cell removal treatment of the membrane biological material is finished, the controller 800 controls the waste liquid removing mechanism to discharge the first waste liquid generated in the cell removal treatment process, the controller 800 controls the cleaning agent liquid storage tank 700 to provide the cleaning agent into the inner tank 200, and the controller 800 controls the rotary driving mechanism 400 to be started to drive the material layering mechanism 300 to rotate, so that the membrane biological material is cleaned in the rotating process along with the material layering mechanism 300;
After the membrane biological material is washed, the controller 800 controls the waste liquid removing mechanism to discharge the second waste liquid generated in the washing process, the controller 800 controls the drying mechanism 103 to dry the inner box 200 after the cell removal treatment, and the controller 800 controls the sterilizing mechanism 104 to sterilize the dried inner box 200.
Compared with the prior art, the cell-removing reaction kettle device carries out cell-removing treatment through an automatic program in the cell-removing treatment process of the membrane biological material, so that the cost of manual liquid adding, liquid changing, cleaning, liquid discharging, drying, sterilizing and the like is greatly reduced, the cell-removing time is saved, the working efficiency is improved, and the beneficial conditions are created for the large-scale production and application of the membrane biological material.
The outer case 100 disclosed in the embodiment of the invention comprises an outer case body, an outer case upper cover 101 arranged on the outer case, and an outer case handle 102 arranged on the outer case upper cover 101, so that the material layering mechanism 300 can be conveniently put in and taken out.
The embodiment of the present invention is not limited to the specific structure of the material layering mechanism 300, and the structure meeting the use requirements of the present invention is within the scope of the present invention.
Referring to fig. 2 and 3, in order to optimize the above embodiment, the material layering device 300 according to the embodiment of the present invention includes at least two layers of storage frames 301 for storing membrane biological materials, wherein any two adjacent layers of storage frames 301 are detachably connected.
Wherein, membrane class biomaterial has all been placed in each layer of storage frame 301, carries out the cell-free processing along with the rotation of material layering mechanism 300 and cell-free solution, so sets up, can avoid membrane class biomaterial to take place the intertwine at cell-free in-process to be located each layered membrane class biomaterial and can fully contact with cell-free reagent through storage frame 301, make cell-free degree more thorough, compare with conventional vibration cell-free processing, can handle membrane class biomaterial in batches simultaneously.
The specific connection mode of the detachable connection is not limited in the embodiment of the invention, the two adjacent layers of storage frames 301 can be connected through the buckle 304, and can be connected through threads, and of course, other connection modes can be adopted, so long as the structure meeting the use requirement of the invention is within the protection scope of the invention.
In order to optimize the above embodiments, the connection manner between the two layers of storage frames 301 disclosed in the embodiments of the present invention is preferably connected by using a buckle 304.
A plurality of through holes 3011 are formed in any one of the storage frames 301, wherein the through holes 3011 communicate the inner cavity of the inner box 200 with the inner cavity of the storage frame 301, and the decellularized reagent or the cleaning reagent entering the inner cavity of the inner box 200 can enter the storage frame 301 through the through holes 3011 to be in contact with the membrane biological material.
The specific arrangement position of the through hole 3011 is not limited in the embodiment of the present invention, and the through hole 3011 may be arranged at the bottom of the storage frame 301, may be arranged at the upper portion of the storage frame 301, or may be arranged at the bottom and the upper portion of the storage frame 301 at the same time, so long as the structure meeting the use requirement of the present invention is within the protection scope of the present invention.
As a preferred embodiment, the through holes 3011 disclosed in the embodiments of the present invention are preferably disposed at the bottom and the upper portion of the storage frame 301 at the same time, so that the decellularization reagent or the cleaning reagent can be fully contacted with the membrane biological material.
In order to avoid leakage of membrane biological materials in the storage frame 301, the storage frame 301 disposed on the top layer of the material layering mechanism 300 disclosed in the embodiment of the present invention is further provided with a storage frame upper cover 302, and of course, the membrane biological materials to be subjected to cell removal treatment can be conveniently placed by the arrangement of the storage frame upper cover 302.
In order to facilitate opening of the upper cover 302 of the storage frame, a storage frame handle 303 is further disposed on the upper cover 302 of the storage frame disclosed in the embodiment of the present invention.
The specific shape of the storage frame 301 is not specifically limited in the embodiment of the present invention, and the structure meeting the use requirement of the present invention is within the protection scope of the present invention.
As a preferred embodiment, the shape of the storage frame 301 disclosed in the embodiment of the present invention preferably adopts a hollow cylindrical structure.
The specifications of the storage frames 301 may be the same or different, so long as the structure meeting the use requirements of the present invention is within the protection scope of the present invention.
As a preferred embodiment, the respective storage frames 301 disclosed in the embodiments of the present invention are preferably set to the same specification, wherein the outer diameter of the storage frame 301 is required to be adapted to the inner diameter of the inner case 200.
The specific dimensions of the inner case 200 and the outer case 100 are not limited in the embodiment of the present invention, and all structures satisfying the use requirements of the present invention are within the scope of the present invention.
The number of specific arrangements of the decellularized reagent liquid storage tank 600 and the cleaning reagent liquid storage tank 700 is not limited in the embodiment of the present invention, and the structures satisfying the use requirements of the present invention are all within the scope of the present invention.
Embodiments of the present invention provide at least one for each of the disclosed decellularization reagent reservoir 600 and cleaning reagent reservoir 700.
Since at least more than two decellularizing reagents are added in the process of decellularizing treatment of heterogeneous materials, the method is used for virus inactivation, immunogenicity removal, degreasing and heat source removal of materials, and in order to avoid mutual interference among different reagents, 4-6 decellularizing reagent liquid storage tanks 600 and 1-2 cleaning reagent liquid storage tanks are preferably arranged.
More preferably, the decellularization reagent reservoir 600 disclosed in the embodiments of the present invention includes a first decellularization reagent reservoir 601, a second decellularization reagent reservoir 602, a third decellularization reagent reservoir 603, a fourth decellularization reagent reservoir 604, and a fifth decellularization reagent reservoir 605.
Wherein, each decellularized reagent liquid storage tank 600 contains different decellularized reagents, so that the arrangement can effectively avoid the influence on the decellularized effect of the membrane biological material due to the impurity of the added solution.
It should be noted that, any one of the decellularized reagent liquid storage tanks 600 is communicated with the inner cavity of the inner tank 200 through the first liquid inlet pipe, and any one of the cleaning reagent liquid storage tanks is communicated with the inner cavity of the inner tank 200 through the second liquid inlet pipe.
In order to better feed the decellularized reagent and the cleaning reagent into the inner cavity of the inner box 200, the decellularized reaction kettle device disclosed by the embodiment of the invention further comprises a liquid inlet 107 which is communicated with the inner cavity of the inner box 200 and the first liquid inlet 606 and the inner cavity of the inner box 200 and the second liquid inlet 107, wherein the first liquid inlet 606 and the second liquid inlet are communicated with the liquid inlet 107 and are communicated with the inner cavity of the inner box 200 through the liquid inlet 107.
It should be noted that, each of the decellularized reagent liquid storage tanks 600 is communicated with the device for providing the decellularized reagent through the first vacuum pump 607, the controller 800 is started, the controller 800 controls the first vacuum pump 607 to be turned on, and under the action of the first vacuum pump 607, the device for providing the decellularized reagent pumps the decellularized reagent into the decellularized reagent liquid storage tank 600, and then enters the inner cavity of the inner tank 200 through the first liquid inlet pipe 606 and the liquid inlet 107.
Of course, each cleaning agent liquid storage tank is communicated with the device for providing the cleaning agent through the second vacuum pump 701, the controller 800 is started, the controller 800 controls the second vacuum pump 701 to be started, and the device for providing the cleaning agent pumps the cleaning agent into the cleaning agent liquid storage tank 700 under the action of the second vacuum pump 701, and then enters the inner cavity of the inner tank 200 through the second liquid inlet pipe and the liquid inlet 107.
The specific structure of the waste liquid removing mechanism is not limited in the embodiment of the invention, and the structure meeting the use requirement of the invention is within the protection scope of the invention.
As a preferred embodiment, the disclosed waste liquid removal mechanism includes a drain 209 in communication with the interior of the inner tank 200, and a drain valve 2091 disposed within the drain 209, wherein the drain valve 2091 is electrically connected to the controller 800. When the decellularization process is completed, the controller 800 controls the drain valve 2091 to be opened, the waste liquid after the decellularization process is discharged through the drain pipe 209, and during the decellularization process, the controller 800 controls the drain valve 2091 to be closed.
Since the membrane type biological material and the solution repeatedly vibrate in the inner box 200 and simultaneously generate friction and frictional heat, the temperature in the inner box 200 is increased, the decellularization treatment effect of the reagent on the membrane type biological material is possibly affected, and even the natural structure of the decellularized membrane type material is possibly damaged, therefore, a temperature control mechanism is necessary to be arranged to reduce the temperature in the inner box 200,
The embodiment of the invention does not limit the specific structure of the temperature control mechanism, and the structure meeting the use requirement of the invention is within the protection scope of the invention.
In order to optimize the above embodiment, the temperature control mechanism disclosed in the embodiment of the present invention includes the cooling pipe 203 disposed on the inner box 200, and the water inlet 204 and the water outlet 205 disposed on the inner box 200, wherein the water inlet 204 and the water outlet 205 are respectively communicated with two ends of the cooling pipe 203, and the cooling water in the cooling pipe 203 can absorb the heat of the inner box 200, thereby reducing the temperature of the inner box 200.
A water inlet valve is disposed at the water inlet 204, and a water outlet valve is disposed at the water outlet 205, wherein both the water inlet valve and the water outlet valve are electrically connected to the controller 800.
When the inner box 200 needs to be cooled, the controller 800 controls the water inlet valve to be opened, and cooling water enters the cooling pipeline 203 through the water inlet 204 to cool the inner box 200, so that the inner box 200 is kept within a preset temperature range, and the membrane biological material is subjected to cell removal treatment within the preset temperature range.
Since the density of water decreases with an increase in temperature, the water having a small density floats above the inner tank 200, and the temperature of the water at a high position in the cooling pipe 203 is high, and in order to improve the circulation cooling efficiency of the cooling water, it is preferable that the water inlet 204 is provided at a lower portion of the inner tank 200, and the water outlet 205 is provided at an upper portion of the inner tank 200, so that the water having a high temperature is preferentially discharged from the cooling pipe 203, thereby improving the cooling efficiency of the cooling water.
In order to facilitate the installation of the cooling duct 203, the side plate of the inner case 200 disclosed in the embodiment of the present invention includes an inner side plate 201 and an outer side plate 202, wherein an accommodating space is provided between the inner side plate 201 and the outer side plate 202, and the cooling duct 203 is disposed in the accommodating space.
In order to effectively prevent heat exchange between the inside and the outside of the inner case 200, the accommodating space disclosed in the embodiment of the present invention is further filled with a thermal insulation material 206, wherein the thermal insulation material 206 is disposed near the inner wall of the outer side plate 202, and the cooling pipeline 203 is disposed near the inner wall of the inner side plate 201. By the arrangement, external heat can be effectively prevented from entering the inner box 200, so that the temperature in the inner box 200 is always kept in a proper range, a proper temperature environment is provided for the decellularization treatment of the membrane biological material, and the effective proceeding of the decellularization treatment is ensured.
When the decellularization of the membrane-based biomaterial is completed, the inner box 200 and the material layering mechanism 300 need to be dried and sterilized, so that a sterile environment is always maintained in the decellularization treatment process of the membrane-based biomaterial.
The embodiment of the present invention does not specifically limit the disclosed drying mechanism 103 and sterilization mechanism 104, as long as the mechanism meeting the use requirements of the present invention is within the scope of the present invention.
In order to optimize the above embodiment, the drying mechanism 103 disclosed in the embodiment of the present invention preferably adopts a circulating fan, wherein one end of the circulating fan is disposed on the outer box 100, the other end of the circulating fan is disposed inside the outer box 100, an air duct 208 is further disposed between the inner side plate 201 and the outer side plate 202 of the inner box 200, and an air inlet 105 and an air outlet 106 are disposed on the outer box 100.
When the decellularization of the membrane biological material is completed and the cleaning of the inner box 200 and the material layering mechanism 300 is completed, the controller 800 controls the circulation fan to be started, the inner box 200 is dried by means of hot air circulation heating and vertical circulation air supply, and hot air enters between the inner side plate 201 and the outer side plate 202 of the inner box 200 from the air duct 208 so as to dry the inner box 200 and the material layering mechanism 300.
Of course, the inner box 200 may also be heated by the electric heating pipe 207, wherein the electric heating pipe 207 is electrically connected to the controller 800, and the controller 800 controls the temperature of the electric heating pipe 207.
Wherein, the air inlet 105 and the air outlet 106 are both disposed on the outer case 100.
When the inner box 200 and the material layering mechanism 300 are required to be sterilized after drying, the sterilization mechanism 104 disclosed in the embodiment of the invention preferably adopts an ultraviolet sterilization lamp, wherein the controller 800 controls the ultraviolet sterilization lamp to be turned on, and the ultraviolet sterilization lamp performs sterilization treatment on the inner box 200 and the material layering mechanism 300.
The reaction kettle device disclosed in the embodiment of the invention further comprises a liquid level sensor 210, wherein the liquid level sensor 210 is electrically connected with the controller, the liquid level sensor 210 can detect the liquid level height change of the inner box 200 in real time and transmit signals to the controller 800, and the controller 800 controls the opening and closing of the first vacuum pump 607 and the second vacuum pump 701 according to the liquid level signals sent by the liquid level sensor 210, so that the decellularized reagent and the cleaning reagent entering the inner box 200 are kept within the preset height range.
In order to effectively support the outer case 100, the cell-free reaction kettle device disclosed in the embodiment of the invention further comprises a supporting mechanism 500 for supporting the outer case 100, wherein the supporting mechanism 500 comprises a hollow supporting case body and rollers 501 arranged at the bottom of the supporting case body, and the rotary driving mechanism 400 is arranged in the supporting case body.
The specific structure of the rotary driving mechanism 400 is not limited in the embodiment of the present invention, and the rotary driving mechanism 400 may be a rotary motor or a rotary cylinder, so long as the structure meeting the use requirement of the present invention is within the protection scope of the present invention.
The rotary driving mechanism 400 disclosed in the embodiment of the present invention preferably employs a rotary motor because of its small size and easy installation.
Wherein, the rotating electrical machines are connected with the bottom of the material layering mechanism 300, in order to facilitate the smooth operation of the rotating electrical machines, bearings 502 are further arranged on the motor shafts of the rotating electrical machines, which not only play a role in supporting the upper cover 101 of the outer box, but also ensure that the rotating electrical machines drive the material layering mechanism 300 to rotate.
The structure of the controller 800 is not specifically limited in the embodiment of the present invention, and the controller 800 may be one or more processors, a memory, a user interface, a network interface, and a communication bus, which are not shown in the figure, but are not specifically limited herein, so long as the programmed control of the decellularization process can be achieved.
The type of the membrane biological material is not limited in the embodiment of the invention, and the membrane biological material can be animal-derived small intestine submucosa, bladder, blood vessel and the like.
Referring to fig. 4, the embodiment of the invention further discloses a decellularizing method, which comprises the following steps:
S100: placing the membrane biological material in the material layering mechanism 300, and controlling the decellularized reagent liquid storage tank 600 to provide the decellularized reagent to the inner tank 200, wherein the decellularized reagent enters the material layering mechanism 300 to be contacted with the membrane biological material;
S200: the rotation driving mechanism 400 is controlled to rotate to drive the material layering mechanism 300 to rotate, and meanwhile, the temperature of the inner box 200 is controlled to be a preset temperature;
S300: when the reaction of the membrane biological material and the decellularized reagent reaches a first preset time, controlling a waste liquid removing mechanism to discharge the waste liquid of the inner box;
s400: controlling the cleaning agent tank 600 to supply the cleaning agent to the inner tank;
S500: controlling the rotary driving mechanism 400 to be started, cleaning the membrane biological materials, and controlling the waste liquid cleaning mechanism to discharge the waste liquid of the inner box after the cleaning reaches a second preset time;
S600: the membrane biological material is taken out from the material layering mechanism 300, the drying mechanism 103 is controlled to dry the inner box, and the sterilization mechanism 104 is controlled to sterilize the dried inner box 200.
When the cell removal treatment is performed on the membrane biological material, the membrane biological material is firstly placed in the material layering mechanism 300 in the inner box 200, then the controller 800 is started, the cell removal reagent is controlled by the controller 800 to be supplied to the inner box 200 from the cell removal reagent liquid storage box 600, the rotation driving mechanism 400 is controlled by the controller 800 to rotate so as to drive the material layering mechanism 300 to rotate, the membrane biological material is fully contacted with the cell removal reagent in the process of rotating along with the material layering mechanism 300, and meanwhile, the temperature of the temperature control mechanism is controlled by the controller 800, so that the temperature of the inner box 200 is kept within a preset temperature range;
After the cell removal treatment of the membrane biological material is finished, the controller 800 controls the waste liquid removing mechanism to discharge the first waste liquid generated in the cell removal treatment process, the controller 800 controls the cleaning agent liquid storage tank 700 to provide the cleaning agent into the inner tank 200, and the controller 800 controls the rotary driving mechanism 400 to be started to drive the material layering mechanism 300 to rotate, so that the membrane biological material is cleaned in the rotating process along with the material layering mechanism 300;
After the membrane biological material is washed, the controller 800 controls the waste liquid removing mechanism to discharge the second waste liquid generated in the washing process, the controller 800 controls the drying mechanism 103 to dry the inner box 200 after the cell removal treatment, and the controller 800 controls the sterilizing mechanism 104 to sterilize the dried inner box 200.
Compared with the prior art, the cell removing method has the advantages that in the cell removing treatment process of the membrane biological material, the cell removing treatment is carried out through an automatic program, so that the cost of manual liquid adding, liquid changing, cleaning, liquid discharging, drying, sterilization and the like is greatly reduced, the cell removing time is saved, the working efficiency is improved, and the beneficial conditions are created for the large-scale production and application of the membrane biological material.
The kind of the cell reagent is not particularly limited in the embodiment of the present invention, and as long as the structure meeting the use requirement of the present invention is within the scope of the present invention, as a preferred embodiment, the embodiment of the present invention includes five decellularization reagents, one cleaning reagent, and of course, a first decellularization reagent liquid storage tank 601, a second decellularization reagent liquid storage tank 602, a third decellularization reagent liquid storage tank 603, a fourth decellularization reagent liquid storage tank 604, a fifth decellularization reagent liquid storage tank 605 and a cleaning reagent liquid storage tank 700 corresponding to the five decellularization reagents are required.
Specifically, the membrane-type biomaterial is placed in the material layering mechanism 300, and then the material layering mechanism 300 is placed in the inner box 200, and the first decellularization reagent is supplied to the inner box 200 by the first decellularization reagent reservoir 601 controlled by the controller 800 for one-time virus inactivation treatment of the membrane-type biomaterial.
Wherein the first decellularization reagent can be a peroxyacetic acid solution or a sodium hydroxide solution, the concentration is 0.1% -2%, and the mass volume ratio (w/v) of the membrane biological material to the decellularization solution is 1:10-1:20, a step of;
The controller 800 is started, and the controller 800 controls the rotation driving mechanism 400 to rotate so as to inactivate viruses on the membrane biological material.
When the first preset time is reached, the controller 800 controls the drain valve 2091 of the waste liquid removing mechanism to open, so as to drain the first waste liquid from the inner tank 200;
The controller 800 controls the cleaning agent tank to supply the cleaning agent to the inner tank 200, wherein the cleaning agent may be phosphate buffer solution or bicarbonate buffer solution, or ultrapure water, and the mass-to-volume ratio (w/v) of the membrane biological material to the cleaning agent is 1:10-1:20, after reaching the second preset time, the controller 800 controls the waste liquid removing mechanism to discharge the second waste liquid generated after the cleaning is completed.
The controller 800 controls the second decellularization reagent tank 602 to supply the second decellularization reagent to the inner tank 200 for removing immunogenicity of the membrane biological material, wherein the second decellularization reagent may be pancreatin solution, peracetic acid solution or sodium hydroxide solution, and has a concentration of 0.1% -2%, and a mass-to-volume ratio (w/v) of the membrane biological material to the decellularization solution is 1:10-1:20, a step of;
the controller 800 rotates the rotation driving mechanism 400 to perform decellularization treatment on the membrane biological material;
after reaching the first preset time, the controller 800 controls the drain valve 2091 of the waste liquid removing mechanism to open, so as to drain the first waste liquid in the inner tank 200;
The controller 800 controls the cleaning agent tank to supply the cleaning agent to the inner tank 200, wherein the cleaning agent may be phosphate buffer solution or bicarbonate buffer solution, or ultrapure water, and the mass-to-volume ratio (w/v) of the membrane biological material to the cleaning agent is 1:10-1:20, after reaching a second preset time, controlling the waste liquid cleaning mechanism to discharge second waste liquid generated after cleaning through the controller 800;
The controller 800 controls the third decellularization reagent tank 603 to supply a third decellularization reagent to the inner tank 200 for degreasing treatment of the membrane biological material, wherein the third decellularization reagent may be a solution mixed with one or more of methanol, chloroform, formaldehyde, methylene chloride, chloroform or diethyl ether, and the mass-to-volume ratio (w/v) of the membrane biological material to the decellularization solution is 1:10-1:20, soaking for 8-20h, and soaking for 1-5 times at normal temperature;
After reaching the first preset time, the controller 800 controls the drain valve 2091 of the waste liquid removing mechanism to open, so as to drain the waste liquid in the inner tank 200;
The controller 800 controls the cleaning agent tank to supply the cleaning agent to the inner tank 200, wherein the cleaning agent may be phosphate buffer solution or bicarbonate buffer solution, or ultrapure water, and the mass-to-volume ratio (w/v) of the membrane biological material to the cleaning agent is 1:10-1:20, after reaching the second preset time, the controller 800 controls the waste liquid removing mechanism to discharge the second waste liquid generated after the cleaning is completed.
The controller 800 controls the fourth decellularization reagent tank 604 to supply a fourth decellularization reagent, which may be sodium dodecyl sulfate, sodium deoxycholate, and Triton X-200 solution, to the inner tank 200 for the heat source removal treatment of the membrane-based biomaterial, with a concentration of 0.1% -0.5%, and a mass-to-volume ratio (w/v) of the membrane-based biomaterial to the decellularization solution of 1:10-1:20, a step of;
the controller 800 rotates the rotation driving mechanism 400 to perform decellularization treatment on the membrane biological material;
After reaching the first preset time, the controller 800 controls the drain valve 2091 of the waste liquid removing mechanism to open, so as to drain the waste liquid in the inner tank 200;
The controller 800 controls the cleaning agent tank to supply the cleaning agent to the inner tank 200, wherein the cleaning agent may be phosphate buffer solution or bicarbonate buffer solution, or ultrapure water, and the mass-to-volume ratio (w/v) of the membrane biological material to the cleaning agent is 1:10-1:20, after reaching the second preset time, the controller 800 controls the waste liquid removing mechanism to discharge the second waste liquid generated after the cleaning is completed.
The controller 800 controls the fifth decellularizing reagent tank 605 to supply the fifth decellularizing reagent to the inner tank 200 for the secondary virus inactivation treatment of the membrane biological material, wherein the fifth decellularizing reagent can be sodium hydroxide or peracetic acid solution with the concentration of 0.1% -5%, and the mass-volume ratio (w/v) of the membrane biological material to the decellularizing solution is 1:10-1:20, a step of;
The controller 800 controls the rotation driving mechanism 400 to rotate, the rotating speed and the time are set, the rotating speed range is 200-300 rpm/min, the time is 1-2h, the temperature is 27-37 ℃, and the cell removal treatment is carried out on the membrane biological material;
after reaching the first preset time, the controller 800 controls the drain valve 2091 of the waste liquid removing mechanism to open, so as to drain the waste liquid in the inner tank 200;
The controller 800 controls the cleaning agent tank to supply the cleaning agent to the inner tank 200, wherein the cleaning agent may be phosphate buffer solution or bicarbonate buffer solution, or ultrapure water, and the mass-to-volume ratio (w/v) of the membrane biological material to the cleaning agent is 1:10-1:20, after reaching the second preset time, the controller 800 controls the waste liquid removing mechanism to discharge the second waste liquid generated after the cleaning is completed.
The rotation speed range of the rotation driving mechanism 400, the first preset time, and the temperature of the inner box 200 are not particularly limited in the embodiment of the present invention, so long as the structure meeting the use requirement of the present invention is within the protection scope of the present invention.
In order to optimize the above embodiment, the rotation speed range of the rotary driving mechanism 400 disclosed in the embodiment of the present invention is preferably 200-300rpm/min, the first preset time is preferably 1-2h, and the temperature of the inner box 200 is preferably 25-40 ℃. ,
In the description of the present application, it should be noted that the directions or positional relationships indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. are based on the directions or positional relationships shown in the drawings, are merely for convenience of describing the present application and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present application. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
Unless specifically stated or limited otherwise, the terms "mounted," "connected," and "coupled" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present application will be understood in specific cases by those of ordinary skill in the art.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (14)
1. The cell-removing reaction kettle device is characterized by comprising an outer box, an inner box, a material layering mechanism, a rotary driving mechanism, a temperature control mechanism, a drying mechanism, a sterilization mechanism, a waste liquid removing mechanism, a cell-removing reagent liquid storage box, a cleaning reagent liquid storage box and a controller;
The inner box is arranged in the outer box, the drying mechanism is arranged in the outer box, the material layering mechanism is arranged in the inner box, and the top of the inner box is of an open structure;
The inner boxes are communicated with a decellularized reagent liquid storage box for providing a decellularized reagent and a cleaning reagent liquid storage box for providing a cleaning reagent, the decellularized reagent liquid storage box is controlled by the controller to supply the decellularized reagent to the inner boxes, the cleaning reagent liquid storage box is controlled by the controller to provide the cleaning reagent to the inner boxes, and the decellularized reagent and the cleaning reagent can enter the material layering mechanism through the inner boxes and are contacted with membrane biological materials in the material layering mechanism; the membrane biological material is animal-derived small intestine submucosa, bladder and blood vessel;
The material layering mechanism comprises at least two layers of storage frames for storing the membrane biological materials, and any two adjacent layers of storage frames are detachably connected; a plurality of through holes are formed in any one of the storage frames, and the through holes are used for communicating the inner cavity of the inner box with the inner cavity of the storage frame;
The rotary driving mechanism is connected with the material layering mechanism and can drive the material layering mechanism to rotate;
The rotary driving mechanism, the drying mechanism, the sterilization mechanism and the waste liquid removing mechanism are all electrically connected with the controller;
When the membrane biological material is subjected to cell removal treatment, the controller controls the cell removal reagent liquid storage tank to supply cell removal reagent to the inner tank, the controller controls the rotation driving mechanism to rotate so as to drive the material layering mechanism to rotate, so that the membrane biological material is fully contacted with the cell removal reagent in the process of rotating along with the material layering mechanism, and meanwhile, the controller controls the temperature of the temperature control mechanism, so that the temperature of the inner tank is kept within a preset temperature range, and the temperature of the inner tank is 25-40 ℃; the temperature control mechanism comprises a cooling pipeline arranged on the inner box, and a water inlet and a water outlet which are arranged on the inner box and are respectively communicated with two ends of the cooling pipeline;
After the membrane biological materials are washed, the membrane biological materials are taken out from the material layering mechanism, the drying mechanism is controlled to dry the inner box, and the sterilizing mechanism is controlled to sterilize the dried inner box.
2. The decellularized reaction kettle device according to claim 1, wherein a storage frame upper cover is further arranged on the storage frame arranged on the top layer of the material layering mechanism.
3. The decellularization reactor device of claim 1, wherein at least one of the decellularization reagent reservoir and the cleaning reagent reservoir is provided;
Any one of the decellularized reagent liquid storage tanks is communicated with the inner cavity of the inner tank through a first liquid inlet pipe;
any one of the cleaning reagent liquid storage tanks is communicated with the inner cavity of the inner tank through a second liquid inlet pipe.
4. The decellularized reaction kettle device according to claim 3, further comprising a liquid inlet which is communicated with the inner cavity of the inner box and the first liquid inlet pipe, and the inner cavity of the inner box and the second liquid inlet pipe, wherein the first liquid inlet pipe and the second liquid inlet pipe are communicated with the liquid inlet and are communicated with the inner cavity of the inner box through the liquid inlet.
5. The decellularized reactor device of claim 1, wherein the waste liquid removal mechanism comprises a drain pipe in communication with the inner cavity of the inner tank, and a drain valve disposed in the drain pipe, the drain valve being electrically connected to the controller.
6. The decellularized reaction kettle device according to claim 1, wherein a water inlet valve is arranged at the water inlet, a water outlet valve is arranged at the water outlet, and the water inlet valve and the water outlet valve are electrically connected with the controller;
the water inlet is arranged at the lower part of the inner box, and the water outlet is arranged at the upper part of the inner box.
7. The decellularized reaction kettle device according to claim 6, wherein the side plate of the inner box comprises an inner side plate and an outer side plate, an accommodating space is arranged between the inner side plate and the outer side plate, and the cooling pipeline is arranged in the accommodating space.
8. The decellularized reaction kettle device according to claim 7, wherein the accommodating space is further filled with a heat insulation material, the heat insulation material is arranged close to the inner wall of the outer side plate, and the cooling pipeline is arranged close to the inner wall of the inner side plate.
9. The decellularized reaction kettle device according to claim 1, wherein the drying mechanism is a circulating fan, one end of the circulating fan is arranged on the outer box, and the other end of the circulating fan is arranged inside the outer box.
10. The decellularized reactor device of claim 1, wherein said sterilization mechanism is an ultraviolet sterilization lamp.
11. The decellularized reactor device of claim 1, further comprising a liquid level sensor disposed in the inner tank, the liquid level sensor being electrically connected to the controller.
12. The decellularized reactor device of claim 1, further comprising a support mechanism for supporting the outer box;
The supporting mechanism comprises a hollow supporting box body and rollers arranged at the bottom of the supporting box body, and the rotary driving mechanism is arranged in the supporting box body.
13. A decellularization method employing the decellularization reaction kettle device of any one of claims 1 to 12, comprising:
S100: placing a membrane biological material in a material layering mechanism, and controlling a decellularized reagent liquid storage tank to provide a decellularized reagent for an inner tank, wherein the decellularized reagent enters the material layering mechanism to be contacted with the membrane biological material;
S200: controlling the rotation driving mechanism to rotate so as to drive the material layering mechanism to rotate, and simultaneously controlling the temperature of the inner box to be a preset temperature;
s300: when the reaction of the membrane biological material and the decellularized reagent reaches a first preset time, controlling a waste liquid removing mechanism to discharge the first waste liquid of the inner box;
S400: controlling a cleaning reagent liquid storage tank to supply cleaning reagent to the inner tank;
s500: controlling the rotary driving mechanism to be started, cleaning the membrane biological materials, and controlling the waste liquid cleaning mechanism to discharge second waste liquid of the inner box after the cleaning reaches a second preset time;
S600: and taking the membrane biological material out of the material layering mechanism, controlling the drying mechanism to dry the inner box, and controlling the sterilizing mechanism to sterilize the dried inner box.
14. The decellularization method of claim 13, wherein the rotational speed of the rotary drive mechanism ranges from 200 to 300rpm/min and the first preset time is 1 to 2 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210231426.2A CN114870772B (en) | 2022-03-10 | 2022-03-10 | Decellularization reaction kettle device and decellularization method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210231426.2A CN114870772B (en) | 2022-03-10 | 2022-03-10 | Decellularization reaction kettle device and decellularization method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114870772A CN114870772A (en) | 2022-08-09 |
| CN114870772B true CN114870772B (en) | 2024-07-12 |
Family
ID=82668429
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210231426.2A Active CN114870772B (en) | 2022-03-10 | 2022-03-10 | Decellularization reaction kettle device and decellularization method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114870772B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114591832B (en) * | 2022-03-28 | 2023-04-18 | 四川大学 | Animal tissue aseptic decellularization device |
| CN118022060B (en) * | 2024-04-12 | 2024-08-13 | 华夏(青岛)生物科技有限公司 | Full-automatic biological tissue decellularization device and method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106267344A (en) * | 2015-05-29 | 2017-01-04 | 深圳艾尼尔角膜工程有限公司 | A kind of cornea takes off cell system |
| CN112830954A (en) * | 2021-01-11 | 2021-05-25 | 西北工业大学 | High-efficiency protein separation and purification method and device based on mixing and deliquoring process |
| CN213273600U (en) * | 2020-09-29 | 2021-05-25 | 江苏优创生物医学科技有限公司 | Electric heating constant-temperature blast drying device for acellular matrix |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6099607A (en) * | 1998-07-22 | 2000-08-08 | Haslebacher; William J. | Rollably positioned, adjustably directable clean air delivery supply assembly, for use in weather protected environments to provide localized clean air, where activities require clean air quality per strict specifications |
| CN100350036C (en) * | 2001-04-27 | 2007-11-21 | 宝洁公司 | Apparatus and method for controlling biofilms |
| JP2009207699A (en) * | 2008-03-04 | 2009-09-17 | Cell Remover:Kk | Decellularization treatment device |
| WO2010056755A2 (en) * | 2008-11-11 | 2010-05-20 | Craig H Randall | Microfluidic embryo and gamete culture systems |
| EP2689438B1 (en) * | 2011-03-23 | 2022-11-16 | Mespilus Inc. | Polarized electrode for flow-through capacitive deionization |
| JP3192933U (en) * | 2014-06-10 | 2014-09-11 | 台灣利得生物科技股▲ふん▼有限公司 | High-efficiency edible bacterial species, cells, or biological material purification equipment |
| CN107708756A (en) * | 2015-04-10 | 2018-02-16 | 哈佛学院院长等 | Immunocyte acquisition equipment and its preparation and application |
| CN107868751A (en) * | 2017-12-07 | 2018-04-03 | 山东隽秀生物科技股份有限公司 | A kind of Full automatic animal tissue takes off cell device and method for removing cells |
| CN210470956U (en) * | 2019-06-06 | 2020-05-08 | 卢氏县天宝生物科技有限公司 | Edible fungus cleaning and drying device |
| CN212494179U (en) * | 2020-05-09 | 2021-02-09 | 宣城市东科电器有限公司 | Capacitor aluminum shell cleaning machine |
-
2022
- 2022-03-10 CN CN202210231426.2A patent/CN114870772B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106267344A (en) * | 2015-05-29 | 2017-01-04 | 深圳艾尼尔角膜工程有限公司 | A kind of cornea takes off cell system |
| CN213273600U (en) * | 2020-09-29 | 2021-05-25 | 江苏优创生物医学科技有限公司 | Electric heating constant-temperature blast drying device for acellular matrix |
| CN112830954A (en) * | 2021-01-11 | 2021-05-25 | 西北工业大学 | High-efficiency protein separation and purification method and device based on mixing and deliquoring process |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114870772A (en) | 2022-08-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114870772B (en) | Decellularization reaction kettle device and decellularization method | |
| CN207325463U (en) | A kind of contagious department medical apparatus cleaning sterilizing device | |
| JP5006860B2 (en) | Desk cleaning device for medical instrument and method for cleaning medical instrument | |
| JP2010115223A5 (en) | ||
| JP4857438B2 (en) | Cleaning sterilizer | |
| CN212199278U (en) | Microbial incubator | |
| CN220049185U (en) | Medical instrument washs drying device | |
| CN216205017U (en) | Textile fabric drying and sterilizing mechanism for textile production workshop | |
| JP5007439B2 (en) | Sterilization apparatus and sterilization method using the same | |
| CN211561207U (en) | Gauze degassing unit for medical care | |
| CN213447389U (en) | Deodorizing device for down feather processing | |
| CN210253289U (en) | High-efficient degassing unit is used to gastroenterology | |
| CN211538684U (en) | Be used for clinical laboratory's equipment degassing unit | |
| CN110037831B (en) | Pig pericardium valve leaf processing apparatus | |
| CN111603583A (en) | Supply room disinfection and sterilization nursing device for preventing nosocomial infection | |
| CN216397354U (en) | Cleaning device for cleaning gynecological speculum | |
| CN221600741U (en) | Multifunctional instrument disinfection device | |
| CN221311655U (en) | Instrument cleaning device for disinfection supply room | |
| CN215124039U (en) | Quick thawing device for food freezing | |
| CN216796118U (en) | A draw blood device for experimental animals and degassing unit and belt cleaning device thereof | |
| CN113458052A (en) | Omnibearing automatic control circulating cleaning device for medical appliances | |
| CN217091472U (en) | Damp-heat sterilization equipment | |
| CN212522489U (en) | Negative pressure isolation ward is with high-efficient sterilization apparatus | |
| CN215194010U (en) | A disinfection belt cleaning device for medical instrument | |
| CN217310195U (en) | Tableware washing and sterilizing equipment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |