TW201928046A - Cell culture module, cell culture system and cell culture method - Google Patents

Abstract

A cell culture module, a cell culture system and a cell culture method are provided. The cell culture system includes a cell tank, a culture medium module and a cell culture module. The cell tank and the culture medium module are communicated to the cell culture module, respectively. The cell culture module includes a case, a first fixer, a second fixer and a sheet of carrier element. The case has a chamber and at least one inlet/outlet. The inlet/outlet is communicated to the chamber. The first fixer is fixed to the case and located in the chamber. The second fixer is disposed in the chamber and is movable relative to the first fixer. The sheet of carrier element is arranged from a plurality of cell culture carriers, and two ends of the sheet of carrier element are fixed to the first fixer and the second fixer, respectively. The sheet of carrier element is in a folded state or an expanded state according to a variation of a distance between the first fixer and the second fixer due to a movement of the second fixer.

Description

Cell culture module, cell culture system and cell culture method

The present disclosure relates to a culture module, a culture system, and a culture method, and more particularly to a cell culture module, a cell culture system, and a cell culture method.

At present, the carrier scaffolds used for cell mass production can be divided into two types, namely natural materials such as collagen, chitosan or gelatin, or synthetic materials such as poly. Ester (PCL), polystyrene (PS), polypropylene (PP) or polylactic acid-polyglycolic acid copolymer (PLGA). Natural materials are mostly derived from animal materials. Although animal materials are low in cytotoxicity and biocompatible, animal materials may carry undetectable animal contamination, so the current trend tends to reduce the use or even the use of animal materials. To reduce the risk of pollution.

In addition, in addition to the related products of alginate as the substrate, the other synthetic materials on the market are quite difficult to degrade, so it is difficult to recover the cells smoothly. Because alginate-based products require high concentrations of calcium ions during cell culture, they may damage cells or cause certain cells to differentiate (eg, mesenchymal stem cells), and calcium is also required for alginate degradation. Ion chelators, which are easily damaged by improper use. In addition, the key technology of vector scaffolds for collecting cells remains to be broken. Therefore, the current cell mass production technology has been stuck in the traditional two-dimensional flat plate culture method, and the process cannot be smoothly scaled up.

Therefore, how to find a carrier material suitable for rapid cell growth without animal pollution sources, and how to improve cell recovery and cell quality is an urgent problem for researchers.

The present disclosure provides a cell culture module, a cell culture system and a cell culture method, which can solve the problem of cell recovery rate and poor cell quality recovery.

The cell culture module of the present disclosure comprises a housing, a first fixing member, a second fixing member and a sheet-shaped carrier member. The housing has a receiving cavity and at least one inlet and outlet. The entrance and exit communicate with the accommodating cavity. The first fixing member is fixed to the housing and located in the accommodating cavity. The second fixing member is disposed in the accommodating cavity and movable relative to the first fixing member. The sheet-shaped carrier is arranged by arranging a plurality of cell culture carriers, and opposite ends of the sheet-shaped carrier are respectively fixed to the first fixing member and the second fixing member. When the distance between the first fixing member and the second fixing member changes with the movement of the second fixing member, the sheet-like carrier member is in an unexpanded state or a folded state.

The cell culture system disclosed herein comprises a cell tank, a culture fluid module and a cell culture module. The cell culture tank and the culture fluid module respectively communicate with the cell culture module, and the cell culture module comprises a shell, a first fixing member, a second fixing member and a sheet-shaped carrier. The housing has a receiving cavity and at least one inlet and outlet. The entrance and exit communicate with the accommodating cavity. The first fixing member is fixed to the housing and located in the accommodating cavity. The second fixing member is disposed in the accommodating cavity and movable relative to the first fixing member. The sheet-shaped carrier is arranged by arranging a plurality of cell culture carriers, and opposite ends of the sheet-shaped carrier are respectively fixed to the first fixing member and the second fixing member. When the distance between the first fixing member and the second fixing member changes with the movement of the second fixing member, the sheet-like carrier member is in an expanded state or a folded state.

The cell culture method of the present disclosure uses the cell culture module described above, the steps of which include attaching the cells to a sheet-like carrier in a folded state. The culture medium is circulated and perfused in a cell culture module, and cell culture is started. The culture solution is drained and the washing solution is poured, and the remaining culture solution is removed by immersion cleaning. The cells are perfused with the de- lysing enzyme, and when the sheet-like carrier is spread, the cells are detached from the sheet-like carrier and suspended in the suspension of the cell culture module. And, recovering a suspension containing cells.

Based on the above, in the cell culture module, the cell culture system, and the cell culture method of the present disclosure, the second fixing member can control the cell culture carrier to change between an untwisted state and a twisted or spread-and-folded state. Improve cell recovery and cell quality.

The above described features and advantages of the present invention will be more apparent from the following description.

The disclosure is more fully described with reference to the drawings of the embodiments. However, the disclosure may be embodied in a variety of different forms and should not be limited to the embodiments described herein. The same or similar reference numerals in the drawings denote the same or similar elements, and the following paragraphs will not be repeated.

1A and FIG. 1B are schematic diagrams showing a twisted state and an untwisted state of a cell culture carrier of a cell culture module according to an embodiment of the present disclosure, respectively. 1A and FIG. 1B, the cell culture module 100A of the present embodiment includes an outer sleeve 110, a first fixing member 120, a second fixing member 130A, and a plurality of cell culture carriers 140. The outer sleeve 110 has a receiving cavity C10 and at least one inlet and outlet T12. The chamber C10 is used to provide a space for cell culture. The inlet and outlet T12 communicates with the accommodating chamber C10. The first fixing member 120 is fixed to the outer sleeve 110 and located in the accommodating cavity C10. The second fixing member 130A is disposed in the accommodating cavity C10 and movable relative to the first fixing member 120. Both ends of the cell culture carrier 140 are respectively fixed to the first fixing member 120 and the second fixing member 130A. When the distance between the first fixing member 120 and the second fixing member 130A changes with the movement of the second fixing member 130A, the cell culture carrier 140 is twisted as shown in FIG. 1A, or the cell culture carrier 140 is untwisted as shown in FIG. 1B. .

In other words, when the distance between the first fixing member 120 and the second fixing member 130A is smaller than the extension length of the cell culture carrier 140 with the movement of the second fixing member 130A, the cell culture carrier 140 is in a twisted state, such as Figure 1A. The cell culture carrier 140 can be strip-shaped. In this state, a plurality of cell culture carriers 140 can be utilized in a limited space to obtain a larger area for cell attachment to increase the number of cells that can be cultured.

In addition, when the distance between the first fixing member 120 and the second fixing member 130A is substantially equal to the extension length of the cell culture carrier 140 as the movement of the second fixing member 130A, the cell culture carrier 140 is in an untwisted state, such as Figure 1B. In this state, the cells can be detached from the cell culture carrier 140 in a process in which the cell culture carrier 140 is switched from the twisted state to the untwisted state by the action of a substance such as an enzyme which can detach the cells from the cell culture carrier 140. Further, since the distance between the cell culture carriers 140 is increased, it is possible to cause an enzyme or the like to act on the inside of the cell culture carrier 140, which is difficult to be sufficiently reacted by the enzyme, thereby contributing to an increase in cell recovery rate.

In this embodiment, since the second fixing member 130A is movably disposed in the accommodating cavity C10, when the outer sleeve 110 is placed in the state of FIG. 1A, the second fixing member 130A may be gravity-dependent. The relationship moves down to a position close to the first fixture 120. When the cell culture carrier 140 is to be converted into an untwisted state, the outer sleeve 110 can be inverted upside down with respect to the state of FIG. 1A as shown in FIG. 1B. As a result, the second fixing member 130A moves downward to a position away from the first fixing member 120 due to its own gravity, and the cell culture carrier 140 is stretched by the first fixing member 120 and the second fixing member 130A. And it is undistorted. Further, in order to enhance the moving ability of the second fixing member 130A, a weight may be attached to the second fixing member 130A to ensure that the second fixing member 130A can move by its own gravity.

From another point of view, when the cell culture carrier 140 is in an untwisted state, each of the cell culture carriers 140 corresponds to a two-dimensional structure. When the cell culture carrier 140 is in a twisted state, the staggered cell culture carrier 140 corresponds to a three-dimensional structure. In FIG. 1A, each of the cell culture carriers 140 in a twisted state is a regular spiral shape, but may also be a disordered twisted state, so that the plurality of twisted state cell culture carriers 140 may exhibit a coil shape, but the present disclosure does not This is limited. In FIG. 1B, each of the cell culture carriers 140 in an undistorted state is straight, so that the plurality of untwisted cell culture carriers 140 may exhibit parallel line arrays, but may also be non-parallel to each other, or a part of the cell culture vector. 140 exhibits a slight bending state, and the disclosure is not limited thereto.

The material of the cell culture carrier 140 is, for example, polyester (Polyester; PET), nylon (Nylon), polyethylene (Polyethylene; PE), polypropylene (PP), polyvinyl chloride (PVC), polystyrene. (polystyrene; PS), polycarbonate (Polycarbonate; PC), ethylene-vinyl acetate copolymer (Ethylene Vinyl Acetate; EVA) or polyurethane (PU). However, the present disclosure is not limited thereto, and any material having a smear characteristic can be used as the material of the cell culture carrier of the present disclosure. Additionally, each of the cell culture carriers 140 may be in the form of strips, linear sheets, or other suitable shapes.

The cell culture carrier 140 can be a cell attachable material or a material that has cell attachability after treatment. The methods of the above treatment include surface modification, surface coating or surface microstructure. The surface modification is, for example, a plasma modification of a surface of a cell attachable material or a cell non-attachable material, and has a cell attachable property on the surface to facilitate cell attachment. Surface coating is applied to a surface of a cell attachable material or a cell attachable material such as collagen, chitosan, gelatin or alginate. Etc., but not limited to this, to facilitate cell attachment. The surface micro-structured is, for example, laser cut on a surface of a cell attachable material or a cell non-attachable material to form micropores to facilitate cell attachment. However, the treatment method of the present disclosure is not limited thereto, and any treatment method capable of improving cell adhesion can be applied to the present disclosure.

The materials of the outer sleeve 110, the first fixing member 120 and the second fixing member 130A are, for example, polyester (Polyester; PET), nylon (Nylon), polyethylene (Polyethylene; PE), polypropylene (Polypropylene; PP), and poly Polyvinyl chloride (PVC), polystyrene (PS), Ethylene Vinyl Acetate (EVA), Polyurethane (PU), Polycarbonate (PC) or Glass Etc., but the disclosure is not limited to this.

In one embodiment, the inlet and outlet T12 of the cell culture module 100 can be disposed at one end of the outer sleeve 110. When the cell culture module 100 is provided with only a single inlet and outlet T12, the medium, the buffer, and the collection of cells are all collected. Liquid can be in and out through the inlet and outlet T12. However, in other embodiments, the overall module is considered to prevent contamination, and the collection of culture medium, buffer, and cells can also be separately accessed through different conduits. In detail, the cell culture module 100 of the present embodiment may include a plurality of inlets and outlets T12, T14, T16, T18 (not shown), T21, T23, wherein the inlet and outlet T12 is disposed at the end of the outer sleeve 110, through The setting of the inlet and outlet T12 allows collection of cells. The inlets and outlets T14, T16 and T18 can be disposed on the side of the outer sleeve 110, adjacent to the end of the outer sleeve 110 provided with the inlet and outlet T12 to provide different buffers and medium inlets, respectively. It should be noted that the number of inlets and outlets T14, T16 and T18 may vary according to the actual type of liquid to be injected, and is not limited to those listed. The inlet and outlet T21 and T23 are disposed at the other end of the outer sleeve 110 relative to the inlet and outlet T12. The inlet and outlet T21 can provide liquid, medium and the like into and out of the inlet and outlet T23, and the inlet and outlet T23 is a reserved hole, wherein the inlet and outlet T21 and the inlet and outlet T14, T16 and T18 are opposite positions. The design helps the distribution and flow of liquid in the receiving chamber C10.

The cell culture module 100 of the present embodiment may further include a spoiler 150 disposed in the accommodating cavity C10 and located between the inlet and outlet T12 and the first fixing member 120. Further, the spoiler 150 can be disposed at the same plane height as the entrances and exits T14, T16 and T18. The liquid entering through the inlet and outlets T14, T16 and T18, through the rotation of the spoiler 150, can drive the liquid in the accommodating chamber C10 to flow, so that the substances in the liquid in the accommodating chamber C10 are evenly distributed.

Cell culture modules can be designed differently depending on whether they are reused or not. When the cell culture module is used in a repetitive manner, the sleeve 110 further includes a body 112 and a cover 114, and the body 112 is coupled to the cover 114 to form a receiving cavity C10. . Through the opening of the lid 114, the chamber C10 can be connected to the accommodation chamber C10 for replacement. In addition, a sealing member 116 may be disposed between the body 112 and the cover 114 to maintain the sealing property of the receiving cavity C10. When the cell culture module is used for one-time use, the outer sleeve 110 is integrally formed.

2A and 2B are schematic diagrams showing the uncultured state and the twisted state of the cell culture carrier of the cell culture module according to another embodiment of the present disclosure. Referring to FIG. 2A and FIG. 2B, the cell culture module 100B of the present embodiment is similar to the cell culture module 100A of FIG. 1A, but it should be noted that the cell culture module 100B of the present embodiment further includes a magnetic control component 160A. . In contrast, the second fixing member 130B of the embodiment has magnetic properties. Therefore, the magnetic control member 160A can control the movement of the second fixing member 130B by a magnetic force such as a magnetic attraction force or a magnetic repulsion force. As shown in FIG. 2A, when the magnetic control member 160A magnetically controls the second fixing member 130B to move to a position away from the first fixing member 120, the cell culture carrier 140 is in an untwisted state. As shown in FIG. 2B, when the magnetic control member 160A controls the second fixing member 130B to move to a position close to the first fixing member 120, the cell culture carrier 140 is in a twisted state. The outer shape of the magnetic control member 160A of the present embodiment substantially matches the outer shape of the outer sleeve 110, and the magnetic control member 160A itself moves, and thereby drives the second fixing member 130B to move, but the disclosure is not limited thereto.

3 is a schematic diagram of a cell culture module in accordance with still another embodiment of the present disclosure. Referring to FIG. 3, the cell culture module 100C of the present embodiment is similar to the cell culture module 100B of FIG. 2A, but it should be noted that the magnetic control member 160B of the present embodiment controls the movement of the second fixing member 130B. The second fixing member 130B is moved to a position away from the first fixing member 120 by magnetic force to directly move to the side of the outer sleeve 110 away from the first fixing member 120. On the contrary, after the magnetic control member 160B is removed, the second fixing member 130B is moved by its own gravity to a position close to the first fixing member 120.

4A and FIG. 4B are schematic diagrams showing a twisted state and an untwisted state of a cell culture carrier of a cell culture module according to still another embodiment of the present disclosure. Referring to FIG. 4A and FIG. 4B, the cell culture module 100D of the present embodiment is similar to the cell culture module 100A of FIG. 1A, but it should be noted that the cell culture module 100D of the present embodiment further includes a fastener 170. In addition, the outer sleeve 110A further has an elastic corrugated structure 110A1. In the unstressed state, the elastic corrugated structure 110A1 is, for example, the extended state of FIG. 4B. The fastener 170 is used to control whether the elastic corrugated structure 110A1 maintains a compressed state or not to switch between the extended state of FIG. 4B and the compressed state of FIG. 4A. The second fixing member 130C of the embodiment is fixed to the outer sleeve 110A, and the elastic crease structure 110A1 is located between the first fixing member 120 and the second fixing member 130C. When the fastener 170 is fastened, the elastic corrugated structure 110A1 is in a compressed state, so that the first fixing member 120 and the second fixing member 130C are close to each other, so that the cell culture carrier 140 is in a twisted state. When the fastener 170 is unfastened, the elastic corrugated structure 110A1 is in an extended state, so that the first fixing member 120 and the second fixing member 130C are apart from each other, so that the cell culture carrier 140 is in an untwisted state. It should be noted that in the embodiment, the fastener 170 is taken as an example, but not limited thereto, and the fastener 170 can be arbitrarily replaced with other fixing components, such as a devil felt, a screw, a rope, and the like.

Figure 5 is a schematic illustration of a cell culture module in accordance with a further embodiment of the present disclosure. Referring to FIG. 5, the cell culture module 100E of the present embodiment is similar to the cell culture module 100A of FIG. 1A, but it should be noted that the cell culture module 100E of the present embodiment further includes a rod member 180 and a guide. Hole 182. The guiding hole 182 is disposed at one end of the outer sleeve 110B relative to the first fixing member 120 to guide the rod member 180 to movably penetrate the outer sleeve 110B. The rod member 180 is coupled to the second fixing member 130A for controlling the movement of the second fixing member 130A. By controlling the extent of the lever member 180 extending into the outer sleeve 110B, the second fixing member 130B can be controlled to move to a position close to or away from the first fixing member 120.

6 is a schematic diagram of a cell culture module in accordance with another embodiment of the present disclosure. Referring to FIG. 6, the cell culture module 100F of the present embodiment is similar to the cell culture module 100A of FIG. 1A, but it should be noted that the cell culture module 100F of the present embodiment further includes a fluid pressure control member 190. The outer sleeve 110 is located in the accommodating cavity C10. The second fixing member 130A is located between the fluid pressure control member 190 and the first fixing member 120. The fluid pressure control member 190 is used to control the movement of the second fixing member 130A. For example, the fluid pressure control member 190 can be applied to a bag body containing a fluid, but the present disclosure is not limited thereto. When the gas, water, oil or other fluid loaded into the fluid pressure control member 190 is increased, the volume of the fluid pressure control member 190 is also increased to push the second fixing member 130A toward the first fixing member 120. mobile. When the gas, water, oil or other fluid loaded into the fluid pressure control member 190 is reduced, the volume of the fluid pressure control member 190 is also reduced, allowing the second fixture 130A to move away from the first fixture 120. mobile.

7 is a schematic diagram of a cell culture module in accordance with still another embodiment of the present disclosure. Referring to FIG. 7, the cell culture module 100G of the present embodiment is similar to the cell culture module 100A of FIG. 1A, but it should be noted that in the cell culture module 100G of the embodiment, the second fixture 130D is screwed to The outer sleeve 110C is on the wall of the hole. In other words, the contact faces of both the second fixing member 130D and the outer sleeve 110C are provided with interfitting threads. Therefore, when the second fixing member 130D rotates relative to the outer sleeve 110C, the second fixing member 130D approaches or moves away from the first fixing member 120. The second fixing member 130D of the embodiment further has a knob 130D1 for facilitating the rotation of the second fixing member 130D by the user.

8A to 8F are schematic views of various stages of cell culture in a cell culture system in accordance with an embodiment of the present disclosure. Referring first to FIG. 8A, the cell culture system 1000 of the present embodiment includes a cell tank 200, a culture fluid module 300, and a cell culture module 400. The cell culture tank 200 and the culture fluid module 300 are respectively connected to the cell culture module 400. The cell culture module 400 may be a cell carrier module of the foregoing embodiments or other cell carrier modules consistent with the spirit of the present disclosure, and a detailed description of the cell culture module 400 is omitted herein. Since the cell culture system 1000 of the present embodiment uses the same cell culture module 400 as the cell carrier module of each of the foregoing embodiments, the cell culture system 1000 of the present embodiment can increase the yield and recovery of cell culture. In addition, the cell culture system 1000 of the present embodiment may optionally further include a pump 500, a cleaning solution tank 600, and a cell desorption enzyme tank 700. The cell tank 200 and the culture fluid module 300 are connected to the cell culture module 400 through the pump 500, respectively. The cleaning solution tank 600 and the cell desorption enzyme tank 700 are also connected to the cell culture module 400, for example, via the pump 500 to the carrier module 400.

The cell tank 200, the culture solution module 300, the cleaning solution tank 600, and the cell desorption enzyme tank 700 of the present embodiment are all connected to the cell culture module 400 through the pump 500. The cell tank 200 is connected to the inlet and outlet T12 of the cell culture module 400 through the pump 500. The cleaning solution tank 600 is connected to the inlet and outlet T14 of the cell culture module 400 through the pump 500. The cell desorbent enzyme tank 700 is connected to the inlet and outlet T16 of the cell culture module 400 through the pump 500. The culture solution module 300 is connected to the inlet and outlet T18 of the cell culture module 400 through the pump 500. The culture solution of the culture solution module 300 enters the cell culture module 400 from the inlet and outlet T18, and then flows back to the culture solution module 300 from the inlet and outlet T21 of the cell culture module 400 at the other end. Therefore, the culture solution of the culture fluid module 300 can be recycled. In order to monitor the quality of the culture solution of the culture fluid module 300, the culture fluid tank 310 of the culture fluid module 300 has a culture fluid sensor 314. The culture fluid sensor 314 is, for example, a pH pH meter, a thermometer, or a dissolved oxygen meter. Further, a stirring rod 312 is disposed in the culture liquid tank 310 to uniformly distribute the culture material in the culture liquid. In addition, the culture fluid module 300 further has a pump 320 and a regulator 330. When the culture liquid sensor 314 senses that the quality of the culture liquid in the culture liquid tank 310 is lower than the critical value, the pump 320 draws the regulator of the regulator 330 into the culture liquid tank 310 to improve the quality of the culture liquid. The cell culture system 1000 of the present embodiment may further include a controller 800. The controller 800 is connected to the cell tank 200, the cleaning solution tank 600, and the cell desorption enzyme through the pump 500, the culture liquid sensor 314, and the pump 320, respectively. The tank 700, the carrier module 400 and the culture fluid module 300 are used to control the pump 500, the pump 320, the regulator 330 and the culture fluid sensor 314. In addition, the cell culture system 1000 of the present embodiment may further include a regulator 900 mounted to the carrier module 400 to adjust the contents of the cell culture module 400 as necessary.

9 is a flow chart of a cell culture method that can be performed by a cell culture system and a cell culture module according to an embodiment of the present disclosure. Referring to FIG. 8A and FIG. 9, when the cell culture is performed, the cell culture carrier 140 of the cell culture module 400 is first twisted, and the setting of each line of the system is completed, step S110. As described in the foregoing embodiments, the method of making the cell culture carrier 140 of the cell culture module 400 in a twisted state is to move the second fixing member to a position close to the first fixing member 120. However, it is not limited to how to perform the method. In the foregoing embodiment, the second fixing member can be displaced by gravity, magnetic force, or other mechanical force. In the embodiment, the second fixing member 130B can be controlled by the magnetic control member 160A. Move, but not limited to this.

Referring to FIG. 8A and FIG. 9 , the cells to be cultured in the cell tank 200 are sent to the cell culture module 400 by the pump 500 to inoculate the cells to be cultured into the cell culture carrier 140 in a twisted state. S120. The cultured cells are, for example, stem cells or differentiated cells, but the disclosure is not limited thereto; specifically, the cultured cells are, for example, VERO, an African green monkey kidney cell line, and adipose stem cells (ADSC, Human Adipose-Derived Stem Cell), Mesenchymal Stem Cell (MSC), Madin-Darby Canine Kidney (MDCK) or Human Embryonic Kidney 293 (HEK293), but Not limited to this. In the present embodiment, the medium is first added to the cell culture carrier 140, and the cells are seeded into the cell culture vector 140. In another embodiment, the cell-containing cell culture medium can also be uniformly added to the cell culture carrier 140. The medium is a standard growth medium commonly used for cell culture, for example, a medium having fetal bovine serum (FBS) or a serum-free medium, but the present disclosure is not limited thereto. In addition, it should be understood that the requirements for the concentration of the cell culture medium are different depending on the characteristics of the cells, so that the concentration of the cells can be adjusted depending on the characteristics of the cells, and growth factors or antibiotics can be added to the medium as the case requires. The cells are attached to a cell culture carrier. In the present embodiment, the cell culture carrier 140 is placed in the carrier module 400 under specific growth conditions (e.g., specific temperature, humidity, or carbon dioxide concentration) to attach the cells to the cell culture carrier 140.

In other embodiments, when the cell culture carrier 140 is in an undistorted state, the setting of each line of the system is first performed, and then the step of inoculating the cells is performed on the undistorted cell culture carrier 140 until the cells are attached. The twisting action of the cell culture carrier 140 is not limited to the above enumeration.

Referring to FIG. 8B and FIG. 9, after the cells are attached, the culture solution is circulated and the cell culture is started, step S130. That is, after the foregoing steps are completed, the pump 500 can be turned on to introduce the culture solution of the culture solution module 300 into the cell culture module 400, and the culture solution is continuously circulated between the culture solution module 300 and the cell culture module 400. To culture cells. The manner of cell culture is, for example, static culture or dynamic culture. The dynamic culture can be carried out by disturbing the culture solution around the cell culture carrier, for example, by using the spoiler 150 as in Fig. 1A, but not shown in the present embodiment. In one embodiment, the number of cells after culture can be increased to more than 100 times the original amount. In another embodiment, the number of cells after culture can be increased to more than 2000 times the original amount.

It should be noted that since different cells have different characteristics, cell culture conditions can be adjusted according to different cell types. For example, when culturing mammalian cells, cell culture can be carried out at 37 ° C and 5% CO ₂ , and the pH of the culture solution is maintained within its physiological range, for example, for most animal cells, culture The pH of the liquid is 7.2~7.4.

Referring to FIG. 8C and FIG. 9, the culture solution is discharged and the cleaning solution is poured, and the remaining culture solution is removed by immersion cleaning, step S140. That is, the culture solution of the cell culture module 400 is returned to the culture solution module 300. Next, the cleaning liquid in the cleaning liquid tank 600 is introduced into the cell culture module 400 by the pump 500, and the remaining culture liquid is removed by immersion cleaning. The cleaning solution is, for example, a phosphate buffered saline solution.

Referring to FIG. 8D and FIG. 9, the cell desorption enzyme is then perfused, and the cell culture carrier and the cells are immersed therein, step S150. That is, the cleaning liquid of the cell culture module 400 is returned to the cleaning solution tank 600. Next, the cell desorption enzyme in the cell desorbing enzyme tank 700 is introduced into the cell culture module 400 by the pump 500, and the cell culture carrier 140 and the cells in a twisted state are immersed therein. The cell desorption enzyme is, for example, trypsin, Tryp LE, Accutase, Accumax or collagenase, but the present disclosure is not limited thereto, and other enzymes or reagents which can desorb cells can also be used.

Referring to FIG. 8E and FIG. 9, the cell culture carrier is then unwrapped to transform into a two-dimensional structure, and the cells are desorbed, step S160. That is, the cell culture carrier 140 of the cell culture module 400 is changed from a twisted state to an untwisted state. As described in the foregoing embodiments, the cell culture carrier 140 of the cell culture module 400 is in an untwisted state by moving the second fixture 130A away from the first fixture 120. In this embodiment, The magnetic control member 160A assists in execution.

Referring to FIG. 8F and FIG. 9, the cell suspension is then recovered, step S170. That is, the cell suspension in the cell culture module 400 is sent to the cell tank 200 through the pump 500. Since the cell culture carrier 140 is subjected to cell recovery in an untwisted state, the loosened structure can sufficiently react the cell culture carrier 140 with the reagent containing the cell de- lysing enzyme, and the loosened structure is also advantageous for growth. The cells in the inner layer of the cell culture carrier 140 are desorbed, thereby effectively increasing the cell recovery rate.

It is to be noted that, in the above embodiment, the cell culture carrier 140 is opened only after the cell desorption enzyme is perfused, and the cells are collected. In other embodiments, however, the cell culture carrier 140 may be first pulled back to a two-dimensional state, and the cell de-enzyme is injected to facilitate collection of the cultured cells, not limited to those enumerated above.

In the above embodiment, the outer sleeve of the cell culture module is cylindrical, but is not limited thereto. In other embodiments, the cell culture module can be designed in other shapes as desired. It should be noted that in order to match the cell culture modules of different shapes, the cell culture carriers are also adaptable and have different laying methods.

10A and FIG. 10B are schematic diagrams showing the sheet-like carrier of the cell culture module in an expanded state and a folded state, respectively, according to an embodiment of the present disclosure. Referring to FIG. 10A and FIG. 10B , the cell culture module 100H of the present embodiment includes a housing 111 , a first fixing member 120 , a second fixing member 130 , and a sheet-shaped carrier 14 . The housing 111 has a receiving cavity C10 and at least one inlet and outlet T12, wherein at least one of the inlets and outlets T12 communicates with the receiving cavity C10. The first fixing member 120 is fixed to the housing 111 and located in the accommodating cavity C10. The second fixing member 130 is disposed in the accommodating cavity C10 and movable relative to the first fixing member 120. The sheet-like carrier 14 is arranged by arranging a plurality of cell culture carriers 140. The opposite ends of the sheet-shaped carrier 14 are respectively fixed to the first fixing member 120 and the second fixing member 130. When the distance between the first fixing member 120 and the second fixing member 130 changes with the movement of the second fixing member 130, the sheet-like carrier member 14 is spread out as shown in FIG. 10A, or the sheet-like carrier member 14 is as shown in FIG. 10B. Shrinking state.

In the present embodiment, the casing 111 is a six-sided box-shaped outer shape, for example, a flat plate shape, but is not limited thereto. When a plurality of cell culture modules 100H are used, the plurality of housings 111 can be stacked with each other to more effectively utilize the entire space. Further, the cell culture carrier 140 is designed to be arranged in a sheet-like carrier 14 in accordance with the change in the outer shape of the housing 111, so that the sheet-like carrier 14 can be covered on one surface of the flat-shaped housing 111 when the sheet-like carrier 14 is in an expanded state.

The straight-line cell culture carrier 140 may be a single-line, double-wire or yarn structure as described above, and the wire may be configured as a straight line, a curve, a spiral curve, a wavy line, a zigzag line, an inverted line or a sheet. Lines, etc., but are not limited to those listed. The plurality of straight-line cell culture carriers 140 are arranged to form a sheet-like carrier 14, and the cell culture carriers 140 may be arranged in parallel or staggered, and the arranged sheet-like carriers 14 may be a single layer or It is multi-layered and is not limited to this.

In the present embodiment, a plurality of cell culture carriers 140 are arranged in a single layer in parallel to form a sheet-like carrier 14, and the cell culture carrier 140 is fixed to the first fixing member 120 and the second at opposite ends of the axial direction. Fixing member 130. Therefore, when the distance between the first fixing member 120 and the second fixing member 130 is smaller than the extending length of the sheet-like carrier 14 (or the cell culture carrier 140) with the movement of the second fixing member 130, the sheet-like carrier 14 (or cell culture carrier 140) will be in a collapsed state, as shown in Figure 10B. In this state, a plurality of cell culture carriers 140 can be utilized in a limited space to obtain a larger area for cell attachment to increase the number of cells that can be cultured. When the distance between the first fixing member 120 and the second fixing member 130 is substantially equal to the extension length of the sheet-like carrier 14 (or the cell culture carrier 140) with the movement of the second fixing member 130, the sheet-like carrier 14 ( Or the cell culture carrier 140) will be in an open state, as shown in Fig. 10A. In this state, the cells can be detached from the cell culture carrier 140 during the transition of the sheet-like carrier 14 from the collapsed state to the expanded state. Further, since the distance between the cell culture carriers 140 is enlarged, a substance such as an enzyme for assisting the detachment of the cells from the cell culture carrier 140 can be reinjected, and it is easy to reach all the cells and sufficiently react, which contributes to Increase cell recovery.

Please refer to Figure 11. In another embodiment, a plurality of cell culture carriers 140 are laid in parallel to form a sheet-like carrier 14, and the sheet-like carrier 14 further includes a first connecting portion 142 and a second connecting portion 144, and cell culture The carrier 140 is fixed to the first connecting portion 142 and the second connecting portion 144 at opposite ends of the axial direction. The first connecting portion 142 and the second connecting portion 144 are adjacently connected to the first fixing member 120 and the second fixing member 130 . In detail, the first connecting portion 142 and the second connecting portion 144 are two long sides of the sheet-like carrier 14 , and the sides of the sheet-shaped carrier 14 that are fixed to the first fixing member 120 and the second fixing member 130 are It is the two short sides of the sheet-like carrier 14. That is, the first connecting portion 142, the second connecting portion 144, the first fixing member 120 and the second fixing member 130 are all located on different sides of the sheet-like carrier 14.

Further, in the embodiment of FIGS. 1A to 10B, the moving direction D of the second fixing member is substantially parallel to the axial extending direction of the cell culture carrier 140, but in the embodiment of FIG. 11, the second fixing member 130 The direction of movement D is substantially perpendicular to the axial extension direction of the cell culture carrier 140 in the sheet-like carrier 14. In other embodiments, the moving direction D of the second fixing member 130 is substantially at an angle to the axial extending direction of the cell culture carrier 140 in the sheet-like carrier 14, and is not limited thereto.

Please refer to FIG. 11 , FIG. 12A and FIG. 12B at the same time. It is to be noted that, in order to simplify the illustration, the cell culture carrier 140 is not illustrated in FIGS. 12A and 12B, but it should be understood from the above description that the configuration of the cell culture carrier 140 can extend the axial direction of the cell culture carrier 140. The direction is perpendicular to the moving direction D of the second fixing member 130, and is angled or parallel. In the embodiment of FIG. 11, when the sheet-like carrier 14 is in the spread state, the first connecting portion 142 and the second connecting portion 144 are substantially straight or slightly curved, but in FIG. 12A and FIG. In the different embodiments of the 12B, the first connecting portion 142 and the second connecting portion 144 can be designed to be zigzag or wavy, and are not limited thereto. The uneven shape of the first connecting portion 142 and the second connecting portion 144, such as a zigzag or a wave shape, can guide the sheet-like carrier 14 to be uniformly folded and compressed when the second fixing member 130 moves toward the first fixing member 120. When the sheet-like carrier 14 is in the folded state, there is a suitable space between the plurality of cell culture carriers 140 to prevent a portion of the region between the plurality of cell culture carriers 140 from being too dense due to uneven extrusion, which is disadvantageous. Cell growth.

As shown in FIG. 12A and FIG. 12B, the sheet-like carrier 14 in the cell culture module 100J further includes a third connecting portion 146. The third connecting portion 146 is disposed between the first connecting portion 142 and the second connecting portion 144 such that one end of the cell culture carrier 140 is fixed to the first connecting portion 142, and the other end of the cell culture carrier 140 extends to the third connecting portion 146. And extending to the second connecting portion 144 via the third connecting portion 146. The third connecting portion 146 has the same shaping function as the folding guide of the first connecting portion 142 and the second connecting portion 144, when the distance between the first connecting portion 142 and the second connecting portion 144 is long, The arrangement of the three connectors 146 provides a fixed support function to prevent the cell culture carrier 140 from sagging due to the distance between the ends being too long. The number of the third connecting portions 146 may not be limited to one.

In an embodiment, the housing 111 has a tapered portion 113, and at least one of the inlets and outlets T12 is disposed at the tapered portion 113. The tapered portion 113 is gradually narrowed outward from the casing 111 to form, for example, a funnel-shaped structure, and the tapered portion 113 allows the cell recovery liquid to easily flow out during cell recovery, thereby reducing cell residue and allowing for accommodation. The gas remaining in the chamber C10 is easily discharged. Of course, the tapered portion 113 can also efficiently collect the liquid entering the accommodating chamber C10 for discharge.

In one embodiment, when the cell culture module is provided with only a single inlet and outlet T12, the medium, the buffer, and the collection of cells, etc., can enter and exit the liquid through the inlet and outlet T12. However, in other embodiments, the overall module is considered to prevent contamination, and the collection of culture medium, buffer, and cells can also be separately accessed through different conduits. In detail, the cell culture module 100H-100J of the present embodiment may include a plurality of inlets and outlets T12, T14, T16 and T18, wherein the inlet and outlet T12 are disposed in the tapered portion 113, and the collection of cells can be performed through the arrangement of the inlet and outlet T12. The inlets and outlets T14, T16 and T18 can be disposed on the other side of the housing 113 relative to the tapered portion 113 to provide different buffer and medium access, respectively. It should be noted that the number of inlets and outlets T14, T16 and T18 may vary according to the actual type of liquid to be injected, and is not limited to those listed. The design of the relative position of the inlet and outlet T12 and the inlet and outlet T14, T16 and T18 facilitates the distribution and circulation of the liquid in the accommodating chamber C10. In addition, the cell culture modules 100H, 100I, and 100J may have different designs depending on whether they are reused or not. When the cell culture modules 100H, 100I, and 100J are used repeatedly, the housing 113 can be designed as two separable components, that is, the housing 113 further includes a body 112 and a cover 114, or a seal can be further disposed. 116 (112, 114, 116 is not shown, refer to FIG. 1A), and its function and purpose are the same as those described above, and are not described herein. When the cell culture modules 100H, 100I, and 100J are disposable, the housing 113 may be integrally formed without being designed in two pieces.

In addition, in the embodiments of the cell culture modules 100H, 100I, and 100J, the second fixing member 130 may be moved by using gravity, magnetic force, or other mechanical force as in the foregoing embodiment, thereby changing the sheet carrier 14 . status. In detail, when gravity is used to displace the second fixing member 130, since the second fixing member 130 is movably disposed in the accommodating cavity C10, when the housing 111 is standing and placed, the second fixing member 130 It moves down to the position close to the first fixing member 120 because of its own gravity. When the sheet-like carrier 14 is to be converted into the unfolded state, the housing 111 can be raised in another corresponding direction, so that the second fixing member 130 moves down to the first position due to its own gravity. The position of the fixing member 120, and the sheet-like carrier member 14 is stretched by the first fixing member 120 and the second fixing member 130 to be in an open state. In addition, if the moving ability of the second fixing member 130 is to be strengthened, a weight may be attached to the second fixing member 130 to ensure that the second fixing member 130 can move by gravity.

Referring to FIG. 13, in an embodiment, when the magnetic force is used to displace the second fixing member 130, the cell culture module 100K further includes a magnetic control member 160K. In contrast, the second fixing member 130K of the embodiment has magnetic properties. Therefore, the magnetic control member 160K can control the movement of the second fixing member 130K by a magnetic force such as a magnetic attraction force or a magnetic repulsion force. When the magnetic control member 160K magnetically controls the second fixing member 130K to move to a position away from the first fixing member 120, the sheet-like carrier member 14 is in an open state. When the magnetic control member 160K controls the second fixing member 130K to move to a position close to the first fixing member 120, the sheet-like carrier member 14 is in a folded state. The magnetic control member 160K of the embodiment is not limited in appearance, as long as it can be close to the surface of the housing 111, and the magnetic control member 160A itself moves, and thereby drives the second fixing member 130K to move, but the disclosure does not Limited to this. In another embodiment, the cell culture module 100K can be placed upright to provide the second fixture 130K to assist in moving displacement due to its own weight.

Referring to FIG. 14 , in an embodiment, the cell culture module 100L further includes a fastener 170 . Further, the housing 111 further has an elastic corrugated structure 111A. The elastic crease structure 111A is the same as the elastic crease structure 110A1 in the cell culture module 100D of FIGS. 4A and 4B described above. In the unstressed state, the elastic crease structure 111A is, for example, the extended state of FIG. The fastener 170 is used to control whether the elastic crease structure 111A maintains a compressed state or not to switch between the extended state and the compressed state. The second fixing member 130L of the embodiment is fixed to the housing 111, and the elastic crease structure 111A is located between the first fixing member 120 and the second fixing member 130L. When the fastener 170 is fastened, the elastic corrugated structure 111A is in a compressed state, so that the first fixing member 120 and the second fixing member 130L are close to each other, so that the sheet-like carrier 14 is in a contracted state. When the fastener 170 is unfastened, the elastic corrugated structure 111A is in an extended state, so that the first fixing member 120 and the second fixing member 130L are apart from each other, so that the sheet-like carrier 14 is in an expanded state. It should be noted that in the embodiment, the fastener 170 is taken as an example, but not limited thereto, and the fastener 170 can be arbitrarily replaced with other fixing components, such as a devil felt, a screw, a rope, and the like.

Referring to FIG. 15, in an embodiment, the cell culture module 100M is similar to the cell culture module 100E of FIG. 5, and further includes a rod member 180 and a guiding hole 182. The guiding hole 182 is disposed at one end of the housing 111 relative to the first fixing member 120 to guide the rod member 180 to be movably disposed through the housing 111. The rod member 180 is coupled to the second fixing member 130 for controlling the movement of the second fixing member 130. By controlling the extent of the lever member 180 extending into the housing 111, the second fixing member 130B can be controlled to move to a position close to or away from the first fixing member 120. It should be noted that, in this embodiment, the second fixing member 130 and the tapered portion 113 are located on different sides of the housing 111 respectively, but in other embodiments, the second fixing member 130 and the tapered portion 113 may be The same side of the housing (ie, the rod 180 and the guiding hole 182 are disposed on the same side as the tapered portion 113) are not limited thereto.

Referring to FIG. 16 , in an embodiment, the cell culture module 100N is similar to the cell culture module 100F of FIG. 6 , and further includes a fluid pressure control member 190 disposed in the housing 111 and located in the receiving cavity C10 . . The second fixing member 130 is located between the fluid pressure control member 190 and the first fixing member 120. The fluid pressure control member 190 is used to control the movement of the second fixing member 130. For example, the fluid pressure control member 190 can be applied to a bag body containing a fluid, but the present disclosure is not limited thereto. When the gas, water, oil or other fluid loaded into the fluid pressure control member 190 increases, the volume of the fluid pressure control member 190 also increases, and the second fixing member 130 can be pushed toward the first fixing member 120. mobile. When the amount of gas, water, oil or other fluid loaded into the fluid pressure control member 190 is reduced, the volume of the fluid pressure control member 190 is also reduced, allowing the second fixing member 130 to move away from the first fixing member 120. mobile. In this embodiment, the second fixing member 130 and the tapered portion 113 are located on different sides of the housing 111 respectively, but in other embodiments, the second fixing member 130 and the tapered portion 113 may be in the same housing. The side (ie, the fluid pressure control member 190 is disposed on the same side as the tapered portion 113) is not limited thereto.

Figure 17 is a schematic illustration of a cell culture system in accordance with an embodiment of the present disclosure. The cell culture system 2000 of the present embodiment includes a cell tank 200, a culture fluid module 300, and a cell culture module 400A. The cell culture tank 200 and the culture fluid module 300 are respectively connected to the cell culture module 400A. The cell culture module 400A may be the cell carrier module (such as the cell carrier module 100H-100N) of the foregoing embodiments or other cell carrier modules conforming to the spirit of the present disclosure, and the details of the cell culture module 400A are omitted herein. Introduction. Since the cell culture system 2000 of the present embodiment uses the same cell culture module 400A as the cell carrier module of each of the foregoing embodiments, the cell culture system 2000 of the present embodiment can increase the yield and recovery of cell culture. In addition, the cell culture system 2000 of the present embodiment may further include a pump 500, a cleaning solution tank 600, a cell desorption enzyme tank 700, and a controller 800. The cell tank 200 and the culture fluid module 300 are connected to the cell culture module 400A through the pump 500, respectively. The cleaning solution tank 600 and the cell desorption enzyme tank 700 are also connected to the cell culture module 400A, for example, via the pump 500 to connect the cell culture module 400. The controller 800 is connected to the cell tank 200, the cleaning solution tank 600, the cell desorption enzyme tank 700, the cell culture module 400A, and the culture fluid module 300 through the pump 500, the culture solution 314, and the pump 320, respectively. The pump 500, the pump 320, the regulator 330, and the culture fluid sensor 314 are controlled.

When performing cell culture, first, the cells to be cultured are attached to the sheet-like carrier 14 in a folded state. In detail, the sheet-like carrier 14 of the cell culture module 400A can be first in a folded state, and the arrangement of the respective lines of the system can be completed. The method of allowing the sheet-like carrier 14 of the cell culture module 400A to be in a folded state is to move the second fixing member 130 to a position close to the first fixing member 120. However, it is not limited to what manner is performed, as described above. In the embodiment, the second fixing member 130 can be displaced by gravity, magnetic force, or other mechanical force, and the details are not described herein. Next, the cells to be cultured in the cell tank 200 are sent to the cell culture module 400A by the pump 500, and the cells to be cultured are inoculated into the sheet-like carrier 14 in a folded state. In other embodiments, when the sheet-like carrier 14 of the cell culture module 400A is in an open state, the pipelines of the system are first set, and then the cells are inoculated and inoculated into the sheet-like carrier in the spread state. On the other hand, the sheet-like carrier 14 of the cell culture module 400A is folded into a folded state to carry out a subsequent cell culture step, which is not limited to the above enumeration.

After the cells are attached, the culture medium is circulated and perfused in the cell culture module 400A, and cell culture is started. Thereafter, the culture solution is drained and the washing solution is poured, and the remaining culture solution is removed by immersion washing.

Next, the cells are desorbed with the enzyme, so that the sheet-like carrier 14 and the cells are immersed therein, and when the sheet-like carrier 14 is spread, the cells are desorbed to the sheet-like carrier 14 and suspended in the cell culture module. 400A suspension. In detail, in one embodiment, the cell desorbing enzyme may be perfused first, and then the sheet-like carrier 14 is unwrapped from a contracted state to an expanded state of a two-dimensional structure. In other embodiments, the sheet-like carrier 14 may first be unwrapped from a collapsed state to an expanded state of a two-dimensional structure, and the cells may be re-perfused with cells, which are not limited to those enumerated.

Finally, the suspension containing the cells is recovered. The detailed operation steps are the same as those of the aforementioned cell culture system 1000, and are not described herein.

As described above, in the cell culture module and the cell culture system of the present embodiment, since the second fixing member is movable, the first fixing member and the second fixing member at both ends of the sheet-shaped carrier are When the distance changes, the sheet-like carrier can be changed between the spread state and the contracted state. The sheet-like carrier in the folded state helps to increase the yield of the cell culture, and the sheet-like carrier in the spread state can improve the cell recovery rate and the quality of the recovered cells.

It should be noted that in the present disclosure, it is mentioned that the cell culture carrier is in an undistorted/twisted state and the sheet-like carrier is in an expanded/retracted state, which essentially means that the cell growth region created by the plurality of cell culture carriers is in the second The change of the dimensional state of the dimension/three-dimensional space is intended to be described in terms of the different laying methods of the cell culture carrier, but is not limited thereto.

The present disclosure has been disclosed in the above embodiments, but it is not intended to limit the disclosure, and any person skilled in the art can make some changes and refinements without departing from the spirit and scope of the disclosure. The scope of protection of this disclosure is subject to the definition of the scope of the appended claims.

100A～100N‧‧‧cell culture module

110, 110A, 110B, 110C‧‧‧ outer sleeve

110A1, 111A‧‧‧ elastic wrinkle structure

111‧‧‧Shell

112‧‧‧Ontology

113‧‧‧attenuation

114‧‧‧ cover

116‧‧‧Seal

120‧‧‧First fixture

130, 130A, 130B, 130C, 130D, 130K, 130L‧‧‧ second fixture

130D1‧‧‧ knob

14‧‧‧Sheet carrier

140‧‧‧ cell culture carrier

142‧‧‧First connection

144‧‧‧Second connection

146‧‧‧ Third connection

150‧‧‧ spoiler

160A, 160B, 160K‧‧‧ magnetic control parts

170‧‧‧fasteners

180‧‧‧ rods

182‧‧‧ Guide hole

190‧‧‧Fluid pressure control

C10‧‧‧容容

D‧‧‧ moving direction

T12, T14, T16, T18, T21, T23‧‧‧ entrances and exits

1000, 2000‧‧‧ cell culture system

200‧‧‧cell slot

300‧‧‧ culture fluid module

310‧‧‧ culture tank

312‧‧‧ stir bar

314‧‧‧ Culture fluid sensor

320, 500‧‧‧

330, 900‧‧‧ adjusters

400, 400A‧‧‧ cell culture module

600‧‧‧cleaning tank

700‧‧‧cell desorption enzyme tank

800‧‧‧ Controller

S110-S170‧‧‧Steps

1A and FIG. 1B are schematic diagrams showing a twisted state and an untwisted state of a cell culture carrier of a cell culture module according to an embodiment of the present disclosure, respectively. 2A and 2B are schematic diagrams showing the uncultured state and the twisted state of the cell culture carrier of the cell culture module according to another embodiment of the present disclosure. 3 is a schematic diagram of a cell culture module in accordance with still another embodiment of the present disclosure. 4A and FIG. 4B are schematic diagrams showing a twisted state and an untwisted state of a cell culture carrier of a cell culture module according to still another embodiment of the present disclosure. Figure 5 is a schematic illustration of a cell culture module in accordance with a further embodiment of the present disclosure. 6 is a schematic diagram of a cell culture module in accordance with another embodiment of the present disclosure. 7 is a schematic diagram of a cell culture module in accordance with still another embodiment of the present disclosure. 8A to 8F are schematic views of various stages of cell culture in a cell culture system in accordance with an embodiment of the present disclosure. 9 is a flow chart of a cell culture method that can be performed by a cell culture system and a cell culture module according to an embodiment of the present disclosure. 10A and FIG. 10B are schematic diagrams showing the sheet-like carrier of the cell culture module in an expanded state and a folded state, respectively, according to an embodiment of the present disclosure. Figure 11 is a schematic illustration of a cell culture module in accordance with another embodiment of the present disclosure. 12A and FIG. 12B are schematic diagrams showing the sheet-like carrier of the cell culture module in an expanded state and a collapsed state, respectively, according to still another embodiment of the present disclosure. Figure 13 is a schematic illustration of a cell culture module in accordance with still another embodiment of the present disclosure. Figure 14 is a schematic illustration of a cell culture module in accordance with a further embodiment of the present disclosure. Figure 15 is a schematic illustration of a cell culture module in accordance with an embodiment of the present disclosure. Figure 16 is a schematic illustration of a cell culture module in accordance with another embodiment of the present disclosure. Figure 17 is a schematic illustration of a cell culture system in accordance with an embodiment of the present disclosure.

Claims (23)

A cell culture module includes: a housing having a receiving cavity and at least one inlet and outlet, wherein the at least one inlet and outlet communicate with the receiving cavity; a first fixing member fixed to the housing and located in the receiving cavity a second fixing member disposed in the accommodating cavity and movable relative to the first fixing member; and a piece-shaped carrier member which is arranged by arranging a plurality of cell culture carriers, the sheet-shaped carrier member The opposite ends are respectively fixed to the first fixing member and the second fixing member, wherein the distance between the first fixing member and the second fixing member changes according to the movement of the second fixing member, the sheet shape The carrier is in an open state or in a collapsed state. The cell culture module according to the first aspect of the invention, wherein the cell culture carriers are respectively fixed to the first fixing member and the second fixing member at opposite ends of the axial extending direction. The cell culture module of claim 1, wherein the sheet-shaped carrier further comprises a first connecting portion and a second connecting portion, wherein opposite ends of the cell culture carriers are respectively fixed to the first a connecting portion and the second connecting portion, and the first connecting portion and the second connecting portion are adjacently connected to the first fixing member and the second fixing member. The cell culture module according to claim 3, wherein the first connecting portion and the second connecting portion are in a zigzag or wavy shape. The cell culture module of claim 3, wherein the sheet-shaped carrier further comprises a third connecting portion disposed between the first connecting portion and the second connecting portion One end of the cell culture carriers is fixed to the first connecting portion, and the other end of the cell culture carriers extends to the third connecting portion, and then extends to the second connecting portion via the third connecting portion. The cell culture module according to claim 1, wherein a moving direction of the second fixing member is perpendicular to an axial extending direction of the cell culture carriers in the sheet-shaped carrier. The cell culture module of claim 1, wherein the housing has a tapered portion, and the at least one inlet and outlet are disposed at the tapered portion. The cell culture module according to claim 1, wherein the number of the at least one inlet and outlet is plural, and is disposed on the tapered portion and the other side of the casing relative to the tapered portion. The cell culture module of claim 1, further comprising a magnetic control member, wherein the second fixing member has magnetic properties, and the magnetic control member is configured to magnetically control the movement of the second fixing member. The cell culture module of claim 1, further comprising a fixing component, wherein the housing further has an elastic corrugated structure for controlling the elastic corrugated structure in an extended state and a The second fixing member is fixed to the housing, and the elastic crease structure is located between the first fixing member and the second fixing member. When the elastic crease structure is in the extended state, the elastic crease structure is in the extended state. When the sheet-shaped carrier is in an open state, and the elastic corrugated structure is in the compressed state, the sheet-like carrier is in a folded state. The cell culture module of claim 1, further comprising a rod movably disposed through the housing and coupled to the second fixing member for controlling movement of the second fixing member. The cell culture module of claim 1, further comprising a fluid pressure control member disposed in the housing and located in the accommodating cavity, wherein the second fixing member is located at the fluid pressure control member The fluid pressure control member controls the movement of the second fixing member between the first fixing members. A cell culture system comprising: a cell culture tank; a culture fluid module; and a cell culture module selected from the group consisting of the cell culture module according to any one of claims 1 to 12, wherein the cell culture tank The culture fluid module is connected to the cell culture module. The cell culture system according to claim 13, further comprising a pump, wherein the cell tank and the culture fluid module respectively communicate with the cell culture module through the pump. The cell culture system according to claim 13 further comprising a cleaning solution tank connected to the cell culture module. The cell culture system according to claim 13 further comprising a cell desorption enzyme tank connected to the cell culture module. The cell culture system of claim 13, further comprising a controller coupled to the cell tank, the culture fluid module, and the cell culture module. A cell culture method according to any one of claims 1 to 12, wherein the method comprises the steps of: attaching a cell to the sheet-like carrier in a contracted state; The culture medium is in the cell culture module, and the culture of the cells is started; the culture solution is drained and a washing solution is poured, and the remaining culture solution is removed by immersion washing; a cell desorbing enzyme is perfused, and the sheet-like carrier is infused In the open state, the cells are detached from the sheet-like carrier and suspended in a suspension of the cell culture module; and the suspension containing the cells is recovered. The cell culture method according to claim 18, wherein the step of attaching the cell to the sheet-shaped carrier in a folded state further comprises: first allowing the sheet-like carrier of the cell culture module to be present In a collapsed state, the cells are then inoculated into the sheet-like carrier in a collapsed state. The cell culture method according to claim 18, wherein the step of attaching the cell to the sheet-like carrier in the contracted state further comprises: inoculating the cell in the spread state of the sheet-like load The sheet-shaped carrier of the cell culture module is then folded into a folded state. The cell culture method according to claim 18, wherein the step of attaching the cell to the sheet-like carrier in a folded state further comprises: moving the second fixing member to be adjacent to the first fixing member The position of the sheet-like carrier of the cell culture module is in a collapsed state. The cell culture method according to claim 18, wherein the cell desorption enzyme is perfused, and when the sheet-like carrier is in an open state, the cell is desorbed to the sheet-like carrier, and suspended in the sheet-like carrier. The step in a suspension of the cell culture module further comprises: first injecting the cell desorbing enzyme, and then converting the sheet-like carrier from a contracted state to an expanded state. The cell culture method according to claim 18, wherein the cell desorption enzyme is perfused, and when the sheet-like carrier is in an open state, the cell is desorbed to the sheet-like carrier, and suspended in the sheet-like carrier. The step of suspending the cell culture module further comprises: first converting the sheet-like carrier from a contracted state to an expanded state, and then perfusing the cell to desorb the enzyme.