CN105483003A - Biological tissue perfusion culture device - Google Patents

Abstract

The invention discloses a biological tissue perfusion culture device. The device can realize the cultivation of biological tissues such as eye tissue, myocardial tissue and bone tissue; and apply centripetal force stimulation, which truly simulates the growth environment of biological tissues and realizes the in vitro cultivation of biological tissues. In the actual application process of the present invention, if the static power supply mode is adopted, the problem of wire winding will be caused. Therefore, the present invention adopts the cooperation of brushes and slip rings to realize dynamic power supply and avoid the problem of wire winding.

Description

A biological tissue perfusion culture device

technical field

The invention relates to the field of tissue culture, in particular to a biological tissue perfusion culture device.

Background technique

With the rapid development of biomedical engineering, there is an urgent need for large-scale artificial culture of cells and biological tissues. At present, the two-dimensional cell culture technology is relatively mature, but the three-dimensional cell culture and tissue-level culture are still very difficult. Especially in the in vitro culture of biological tissue, how to make nutrients enter the tissue to avoid the phenomenon of "central necrosis" is still an urgent technical problem to be solved. On the other hand, there is a wide range of motion and deformation in the living body, which has a key impact on the growth and decline of biological tissues. Therefore, how to apply mechanical stimuli, especially physical force (centripetal force) stimuli that simulate gravity in in vitro tissue culture is also of great importance. Technology with important application value. At present, NASA's Johnson Space Center has developed a rotating wall bioreactor (RWV), and on this basis, it has developed a cell rotation culture system (RCCS), which can make suspension cells subject to The gravity vector is zero. However, this type of experimental device is mainly used for in vitro culture at the cell level, and it is not easy to achieve in vitro culture at the biological tissue level, so designing a device suitable for biological tissue culture has important application significance.

Contents of the invention

In view of this, the present invention provides a biological tissue perfusion culture device, which realizes in vitro culture of biological tissue by loading the centripetal force provided by the tissue.

A biological tissue perfusion culture device, including a perfusion component, a loading component and a fixing component;

The perfusion assembly is fixed to the loading assembly, and the loading assembly is used to carry the perfusion assembly; the fixing assembly is fixed to the loading assembly to provide fixed support for the loading assembly;

The perfusion component is used to provide the required growth environment for biological tissue;

The loading component is used to apply centripetal force to the perfusion component.

Preferably, the perfusion assembly includes a perfusion chamber, a peristaltic pump, a stepper motor drive controller, a first brush, a second brush, a slip ring and a transfer bottle;

Wherein, the slip ring mainly includes two copper rings;

The perfusion cavity, the peristaltic pump and the transfer bottle are sequentially connected end to end through rubber tubes to form a loop; the stepper motor drive controller is connected to the peristaltic pump; the first brush and the second brush are connected to the stepper in the perfusion component through wires. The motor drive controller is connected; the first brush is in contact with one of the copper rings; the second brush is in contact with the other copper ring to realize current conduction.

The loading assembly includes: a main shaft, a peristaltic pump fixing device, a transfer bottle fixing device, a bearing and a shaft sleeve;

The main shaft and the bearing are installed and fixed on the base of the fixed assembly, and are perpendicular to the upper surface of the base; the shaft sleeve is sleeved with the main shaft through its central hole and fixed on the upper surface of the base; the transfer bottle fixing device and the peristaltic pump fixing device The through holes pass through the main shaft in turn, are socketed on the outer surface of the main shaft and fixed;

The fixing component is a fixing frame with a base, a bracket and a top plate.

Its connection relationship is:

The slip ring passes through the main shaft and is fixed on the upper surface of the base, and the inner diameter of the slip ring is larger than the outer diameter of the shaft sleeve. At different positions on the lower surface of the device; the transfer bottle is placed on the transfer bottle fixing device; the peristaltic pump and the stepper motor drive controller connected to the peristaltic pump are placed on the peristaltic pump fixing device; the perfusion chamber is fixed on the main shaft;

A motor is housed on the top plate of the fixed mount, and the motor cooperates with the main shaft.

Preferably, the structures of the peristaltic pump fixing device and the transfer bottle fixing device are as follows:

The peristaltic pump fixing device is a disc structure, and the center of the disc is provided with a through hole for connecting with the main shaft; along the outer circumference of the disc, there are several large openings for hanging the peristaltic pump; between two adjacent large openings There is a small opening between;

The transfer bottle fixing device is made of nylon material; the transfer bottle fixing device is provided with a bottle holder and a bottle clamp; the bottle holder is a disc structure, the center of the bottle holder is provided with a through hole for connecting with the main shaft, The upper end of the holder is provided with a plurality of bottle clamps along the outer circumference of the bottle holder; a plurality of openings are arranged along the outer circumference of the bottle holder; a plurality of threaded holes for fixing with electric brushes are arranged on the bottle holder;

Preferably, the opening of the transfer bottle fixing device fixed on the main shaft communicates with the large opening of the peristaltic pump fixing device.

Preferably, the perfusion cavity in the perfusion assembly is a solid structure; centrally symmetrical installation holes are provided on the perfusion cavity; the perfusion cavity is divided into three sections, and the middle section is used to place biological tissues The two sides of the sample are respectively threaded sections, and the inner diameter of the threaded section is larger than the inner diameter of the middle section; one side of the perfusion cavity is also provided with a threaded hole on the end surface.

Preferably, the sealing bolt is a stepped cylinder, which is divided into two sections; wherein, the large-diameter section is provided with external threads, and the outer surface of the small-diameter section is provided with barbs.

Preferably, the perfusion assembly further includes a flange assembly; the loading assembly further includes a cantilever for fixing the flange assembly to the main shaft;

The flange assembly is an integral structure composed of a flange chassis and a cylindrical barrel; the cylindrical barrel is located above the flange chassis; and the cylindrical barrel is provided with openings along the axial direction, and the rest of the cylinder is uniformly provided with through holes;

The cantilever has the same structure as the cylinder in the flange assembly, and the outer diameter of the cantilever is equal to the inner diameter of the cylinder;

The flange assembly is fixed on one side of the perfusion cavity, and the cantilever is correspondingly nested on the inner surface of the cylinder; wherein, the correspondence here means that the opening of the cantilever and the opening of the flange cylinder are located on the same side and communicate with the outside world; The connecting surface of the cantilever and the main shaft is set as an arc structure matching the outer peripheral surface of the main shaft.

Beneficial effect:

1. Using this device, the cultivation of biological tissues such as ocular tissue, myocardial tissue and bone tissue can be realized; and the centripetal force stimulation can be applied to truly simulate the growth environment of biological tissues, and the in vitro cultivation of biological tissues can be realized.

2. In the actual application process of the present invention, if the static power supply method is adopted, the problem of wire winding will be caused. Therefore, the present invention adopts the cooperation of brushes and slip rings to realize dynamic power supply and avoid the problem of wire winding .

3. In order to realize the use of more peristaltic pumps and transfer bottles in the experimental process, the present invention designs the peristaltic pump fixing device and the transfer bottle fixing device with multiple openings for fixing the peristaltic pumps and transfer bottles, so as to achieve limited In the space available, more peristaltic pumps and transfer bottles are used.

4. In the present invention, multiple openings are provided on the disk of the peristaltic pump fixing device and the bottle holder of the transfer bottle fixing device. During installation, different openings are installed in a corresponding or staggered manner to reduce their axial height , making the device compact.

5. The perfusion cavity with this structure can have a good drainage effect on the passing nutrient solution, ensuring that the biological tissue can fully absorb nutrients.

6. The barbs on the sealing bolt can be well fixed with the rubber tube.

7. In order to fix the perfusion cavity on the main shaft more stably, the present invention designs a flange assembly and a cantilever, and through the cooperation of the two, the perfusion cavity is fixed on the main shaft.

Description of drawings

Figure 1 is an assembly diagram of the perfusion assembly.

Figure 2 is an assembly diagram of the loading assembly.

Figure 3 is an assembly diagram of the fixing components.

Fig. 4 is a sectional view of the sealing bolt.

Figure 5(a) is a three-dimensional view of the flange assembly.

Figure 5(b) is a cross-sectional view of the flange assembly.

Fig. 6 is a sectional view of the slip ring.

Fig. 7(a) is a three-dimensional view of the first brush.

Fig. 7(b) is a sectional view of the first brush.

Fig. 8(a) is a three-dimensional view of the second brush.

Fig. 8(b) is a sectional view of the second brush.

Fig. 9 is a cross-sectional view of the perfusion cavity.

Figure 10(a) is a three-dimensional view of the cantilever.

Figure 10(b) is a cross-sectional view of the cantilever.

Fig. 11 is a top view of the peristaltic pump fixing device.

Fig. 12(a) is a three-dimensional view of the transfer bottle fixing device.

Figure 12(b) is a top view of the transfer bottle fixing device.

Figure 13 is a three-dimensional view of the shaft sleeve.

Figure 14(a) is the overall structure diagram.

Figure 14(b) is the overall assembly diagram.

Among them, 1-flange assembly, 2-perfusion cavity, 3-O-ring, 4-sealing bolt, 5-rubber tube, 6-peristaltic pump, 7-transfer bottle, 8-stepping motor drive controller, 91 -First brush, 92-second brush, 10-slip ring, 11-flange chassis, 12-flange cylinder, 13-main shaft, 14-cantilever, 15-motor, 16-shaft sleeve, 17- Large opening, 18-small opening, 19-bottle holder, 20-bottle clamp, 21-peristaltic pump fixing device, 22-transfer bottle fixing device, 23-bearing, 24-top plate, 25-bracket, 26-base, 27- Springs, 28-carbon rods.

detailed description

The present invention will be described in detail below with reference to the accompanying drawings and examples.

The invention provides a biological tissue perfusion culture device, comprising a perfusion component, a loading component and a fixing component;

As shown in Figure 1 and Figure 2, the perfusion assembly includes: rubber tube 5, peristaltic pump 6, transfer bottle 7, perfusion cavity 2, O-ring 3, sealing bolt 4, flange assembly 1, stepping motor drive Controller 8, first brush 91, second brush 92 and slip ring 10;

As shown in Figure 4; the sealing bolt is a stepped cylinder, which is divided into two sections; wherein, the large diameter section is provided with external threads, and the outer surface of the small diameter section is provided with barbs; in order to avoid excessive extrusion of the O-ring, On the step surface formed by the large-diameter section and the small-diameter section, a boss is epitaxially formed;

As shown in Fig. 5 (a) and Fig. 5 (b); Described flange assembly is the integral structure that flange chassis 11 and cylinder 12 form; Cylindrical barrel 12 is positioned at the top of flange chassis 11; And cylindrical barrel 12 Openings are provided along the axial direction, and through holes are uniformly provided in the rest;

As shown in Figure 6; the slip ring includes a copper ring 27, an insulating ring 28 and an insulating base 29; the insulating base is a cylindrical structure, and one end of the cylindrical tube extends inward to form a boss; two copper rings are embedded in the cylindrical tube On the inner circumference of the inner circumference, the insulating ring is located between two copper rings;

Both the first electric brush 91 and the second electric brush 92 are rectangular parallelepiped; one side end surface of the rectangular parallelepiped is provided with a through hole; a spring is arranged in the through hole; one end of the spring is in conflict with the bottom surface of the through hole, and the other end of the spring is in contact with The carbon rods interfere; and the carbon rods in the first electric brush 91 are in contact with one of the copper rings; the carbon rods in the second electric brush 92 are in contact with another copper ring; as shown in Fig. 7 (a), Fig. 7 (b), Shown in Fig. 8 (a) and Fig. 8 (b); In order to reduce the mass of electric brush, the present invention makes first electric brush 91 and second electric brush 92 lower surfaces into arc surface, and unsupported in electric brush Removal of the stressed part;

As shown in Figure 9; the perfusion cavity is a solid structure. In this embodiment, a cylindrical structure is adopted; the perfusion cavity is provided with centrally symmetrical mounting holes; the perfusion cavity is divided into three-stage structures , the middle section is used to place biological tissue samples such as eye tissue, myocardial tissue and bone tissue, and the two sides are respectively threaded sections, and the inner diameter of the threaded section is larger than the inner diameter of the middle section. A threaded hole is also provided on the end surface of one side of the perfusion cavity;

The connection relationship is that: the perfusion components constitute two major circuits; they are liquid circuits composed of O-rings, sealing bolts, perfusion chambers, rubber tubes, peristaltic pumps and transfer bottles; they are driven and controlled by peristaltic pumps and stepping motors. A current loop composed of a switch, a brush and a slip ring;

Liquid circuit: the O-rings 3 are respectively placed on the stepped surface formed by the two threaded sections and the middle section of the perfusion cavity 2; solid together. On the perfusion cavity, the flange assembly 1 is fitted on the outer surface of the sealing bolt on the side with the threaded hole, and is fixedly connected with the perfusion cavity after being threaded through the threaded hole; the small diameter section of the sealing bolt is connected with a rubber tube, and the rubber tube After passing through the central hole of the flange base and the opening of the cylinder, it is connected with the peristaltic pump 6 and the transfer bottle 7; the small diameter section of the sealing bolt at the other end of the perfusion cavity is connected with the transfer bottle; a fluid circuit is formed.

Current loop: the peristaltic pump 6 is also connected with the stepper motor drive controller 8 and two brushes through wires in turn; the brushes form contact with the slip ring to realize the conduction of the current.

As shown in Figure 2; the loading assembly includes: a main shaft 13, a cantilever 14, a peristaltic pump fixing device 21, a transfer bottle fixing device 22, a bearing 23 and a shaft sleeve 16;

As shown in Figure 10(a) and Figure 10(b); the cantilever has the same structure as the cylinder 12 in the flange assembly, and the outer diameter of the cantilever is equal to the inner diameter of the cylinder; correspondingly nested in the cylinder The inner surface of 12; the cantilever is used to realize the connection between the flange assembly and the main shaft; in order to better realize the connection between the cantilever and the main shaft, the end surface of the non-open end of the cantilever is set as an arc structure matching the outer circumferential surface of the main shaft;

As shown in Figure 11; the peristaltic pump fixing device 21 is a disc structure, and the center of the disc is provided with a through hole for connecting with the main shaft; there are a plurality of large holes for hanging the peristaltic pump along the outer circumference of the disc. An opening 17; a small opening 18 is arranged between two adjacent large openings;

As shown in Figure 12; the transfer bottle fixing device 22 is made of nylon material; the transfer bottle fixing device is provided with a bottle holder 19 and a bottle clamp 20; the bottle holder is a disc structure, and the center position of the bottle holder is provided with a In the through hole connected with the main shaft, the upper end of the bottle holder is provided with a plurality of bottle clips along the outer circumference of the bottle holder; in order to make better use of the space, a plurality of openings are provided along the outer circumference of the bottle holder; There are multiple threaded holes on the bracket for fixing with the brush;

As shown in Figure 13; the shaft sleeve 16 is a flange structure;

The connection relationship is that a bearing 23 is embedded inside the base 26 in the fixed assembly, the main shaft 13 is installed on the inner ring of the bearing 23, and is perpendicular to the upper surface of the base 26; the shaft sleeve 16 passes through the center hole of the flange Socketed with the main shaft 13 and fixed on the upper surface of the base 26; the transfer bottle fixing device 22 and the peristaltic pump fixing device 21 are sequentially connected with the main shaft through through holes and fixed with the main shaft 13; the cantilever 14 is located above the peristaltic pump fixing device 21, And welded on the main shaft 13; in order to reduce the axial dimension of the whole system, the present invention needs to correspond the opening in the transfer bottle fixing device 22 to the large opening 17 of the peristaltic pump fixing device 21;

As shown in Figure 3; the fixed assembly is a fixed frame with a base 26, a support 25 and a top plate 24;

As shown in Figure 14 (a) and (b); the connection relationship between the perfusion assembly, the loading assembly and the fixing assembly is: the slip ring 10 passes through the main shaft 13 and is fixed on the upper surface of the base 26, and the inner diameter of the slip ring greater than the outer diameter of the shaft sleeve, the shaft sleeve and the slip ring form a nested structure; the first brush 91 and the second brush 92 are respectively installed on the lower surfaces of two different bottle holders 19; and the first brush The carbon rod in 91 is in conflict with the copper ring 27 above; the carbon rod in the second brush 92 is in conflict with the copper ring 27 below; the cylinder in the flange assembly is set on the outer surface of the cantilever 14, and the opening of the cantilever It is located on the same side as the opening of the flange cylinder and communicates with the outside world; the transfer bottle in the perfusion assembly is placed on the bottle holder, and the transfer bottle body is stuck in the bottle clamp; the pump head of the peristaltic pump is placed in the large opening, The stepper motor drive controller 8 connected with the peristaltic pump is located directly above the opening in the transfer bottle fixture 22;

The rubber tube between the perfusion chamber and the peristaltic pump is connected through the opening of the cantilever and the flange assembly; the rubber tube between the peristaltic pump and the transfer bottle passes through the small opening 18 provided by the peristaltic pump fixing device 21;

A motor 15 is housed on the top plate 24 of the fixed mount, and the motor 15 cooperates with the main shaft 13;

To sum up, the above are only preferred embodiments of the present invention, and are not intended to limit the protection scope of the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims (7)

1. A biological tissue perfusion culture device, characterized in that: comprising a perfusion assembly, a loading assembly and a fixing assembly; The perfusion assembly is fixed to the loading assembly, and the loading assembly is used to carry the perfusion assembly; the fixing assembly is fixed to the loading assembly to provide fixed support for the loading assembly; The perfusion component is used to provide the required growth environment for biological tissue; The loading component is used to apply centripetal force to the perfusion component. 2. biological tissue perfusion culture device as claimed in claim 1, is characterized in that: The perfusion assembly includes a perfusion chamber (2), a peristaltic pump (6), a stepper motor drive controller (8), a first brush (91), a second brush (92), a slip ring (10) and Transfer bottle (7); Wherein, the slip ring (10) mainly includes two copper rings; The perfusion cavity (2), the peristaltic pump (6) and the transfer bottle (7) are sequentially connected head to tail through the rubber tube (5) to form a circuit; the stepper motor drive controller (8) is connected to the peristaltic pump (6); The first electric brush (91) and the second electric brush (92) are connected with the stepper motor drive controller (8) in the perfusion assembly through wires; the first electric brush (91) forms contact with one of the copper rings; The second electric brush (92) is in contact with another copper ring to realize the conduction of electric current. The loading assembly includes: a main shaft (13), a peristaltic pump fixing device (21), a transfer bottle fixing device (22), a bearing (23) and a shaft sleeve (16); The installation of the main shaft (13) and the bearing (23) is fixed on the base (26) of the fixed assembly, and is perpendicular to the upper surface of the base (26); Socketed and fixed on the upper surface of the base (26); the through holes of the transfer bottle fixing device (22) and the peristaltic pump fixing device (21) pass through the main shaft (13) in sequence, and are sleeved on the outer surface of the main shaft and fixed; The fixing assembly is a fixing frame with a base (26), a support (25) and a top plate (24). Its connection relationship is: The slip ring (10) passes through the main shaft (13) and is fixed on the upper surface of the base (26), and the inner diameter of the slip ring (10) is larger than the outer diameter of the shaft sleeve (16), and the shaft sleeve (16) and the slip ring (10) into a nested structure inside and outside; the first electric brush (91) and the second electric brush (92) are installed at different positions on the lower surface of the transfer bottle fixing device (22) respectively; the transfer bottle (7) is placed in the transfer bottle fixing device ( 22); the peristaltic pump (6) and the stepper motor drive controller (8) connected to the peristaltic pump (6) are placed on the peristaltic pump fixing device (21); the perfusion chamber (2) is fixed on the main shaft; Motor (15) is housed on the top plate (24) of fixed mount, and described motor (15) cooperates with main shaft (13). 3. biological tissue perfusion culture device as claimed in claim 2, is characterized in that: described peristaltic pump fixture (21) and transfer bottle fixture (22) structure is as follows: The peristaltic pump fixing device (21) is a disc structure, and the center of the disc is provided with a through hole for connecting with the main shaft (13); there are a plurality of large holes for hanging the peristaltic pump (6) along the outer circumference of the disc. An opening (17); a small opening (18) is arranged between two adjacent large openings; The transfer bottle fixing device (22) is made of nylon material; the transfer bottle fixing device is provided with a bottle holder (19) and a bottle clamp (20); the bottle holder is a disc structure, and the center of the bottle holder is provided with a In the through hole connected with the main shaft (13), a plurality of bottle clamps are arranged on the upper surface of the bottle holder along the outer circumference of the bottle holder; a plurality of openings are arranged along the outer circumference of the bottle holder; a threaded hole for fixing with the brush; 4. The biological tissue perfusion culture device according to claim 3, characterized in that: the opening of the transfer bottle fixing device (22) fixed on the main shaft communicates with the large opening (17) of the peristaltic pump fixing device (21). 5. biological tissue perfusion culture device as claimed in claim 2, is characterized in that: the perfusion chamber body (2) in the described perfusion assembly is a solid structure; The perfusion chamber body is provided with center-symmetric installation holes; The perfusion cavity is divided into three sections, the middle section is used to place biological tissue samples, and the two sides are respectively threaded sections, and the inner diameter of the threaded section is larger than the inner diameter of the middle section; the end surface of one side of the perfusion cavity is There are also threaded holes. 6. The biological tissue perfusion culture device as claimed in claim 2, characterized in that: the sealing bolt (4) is a stepped cylinder, divided into two sections; wherein, the large diameter section is provided with external threads, and the small diameter section There are barbs on the outer surface. 7. The biological tissue perfusion culture device according to claim 2, characterized in that: the perfusion assembly also includes a flange assembly (1); the loading assembly also includes a flange assembly (1) for fixing the main shaft the cantilever (14); The flange assembly is an integral structure composed of a flange chassis (11) and a cylindrical barrel (12); the cylindrical barrel (12) is located above the flange chassis (11); and the cylindrical barrel (12) is provided with an opening along the axial direction, The remaining parts are uniformly provided with through holes; The cantilever has the same structure as the cylinder (12) in the flange assembly, and the outer diameter of the cantilever is equal to the inner diameter of the cylinder; The flange assembly is fixed on one side of the perfusion chamber (2), and the cantilever is correspondingly nested on the inner surface of the cylinder (12); wherein, the correspondence here means that the opening of the cantilever and the opening of the flange cylinder are located on the same side , and communicate with the outside world; the connecting surface of the cantilever and the main shaft is set as an arc-shaped structure matching the outer peripheral surface of the main shaft.