Cell damage repair incubator and its application

Abstract

The present invention belongs to medical instruments, and is a cell damage repair incubator and its application, which is composed of a support and fixation system A, a conditioning and culture system B, a filtration and circulation system C, and an oxygen supply and ventilation system D. Its main features and advantages are as follows: ① It is convenient to install and disassemble between systems and within different systems, which is beneficial for cleaning and sterilization; ② The stirring is gentle and uniform, without damaging the cultured cells; ③ The unique design not only reduces contamination, but also avoids the adhesion of the probe part of the cultured cells immersed in the culture medium, simplifies the operation and reduces the cost; ④ The temperature change is gentle and does not damage the cells; ⑤ The built-in universal rotating shaft is beneficial for preventing the cultured cells from clogging the conical filter screen; ⑥ It has good hydrodynamic advantages and avoids the retention of cultured cells at the bottom; ⑦ It is easy to operate, inexpensive, and convenient for popularization and application. It can be applied to the repair of damaged cells, the recovery after poisoning, and the adjustment of sub-healthy states for treatment, production, and research purposes.

Description

Technical Field

[0001] The present invention belongs to medical instruments, and particularly relates to a cell damage repair incubator, which can be applied to the repair of damaged cells, the recovery after poisoning, and the adjustment of sub-healthy states for treatment, production, and research purposes. Background Art

[0002] Many primary and subcultured cells of humans and animals are widely used in the biomedical field. For example, stem cells and immune cells are used to treat diseases; genetically engineered cells are used to produce protein drugs; BHK and Vero cells are used to produce vaccines; seed cells for tissue engineering; cells for scientific research, etc.

[0003] However, these cells, which are very important in the fields of clinical, production, and scientific research, often have certain defects. For example, during the processes of cell isolation and preparation, digestion and passage, cryopreservation and resuscitation, etc., cell damage is inevitable; the use of reagents or drugs in the source organism and in the processes of cell isolation, culture, passage, cryopreservation, etc. may cause cell poisoning; due to reasons such as cell isolation and preparation, cryopreservation and resuscitation, etc., the sub-healthy state of cells may occur. These problems of cells need to be detected by professional means and are often ignored by people.

[0004] Since human and animal cells do not have the protection of cell walls like plants, and their growth is relatively slow, defects such as cell damage, poisoning, and sub-healthy state are not easily self-repaired, detoxified, or restored. In order to ensure the quality of human and animal cells, it is necessary to repair, detoxify, and recover the cells with defects such as damage, poisoning, and sub-healthy state through culture, so as to obtain better application effects in treatment, production, and research. In particular, for therapeutic cells used in clinical transplantation, since damaged cells are more likely to die or apoptose in the body, it directly affects the effect of cell therapy and may even lead to medical risks.

[0005] Currently, specialized cell repair culture instruments are still very lacking and there is an urgent need for research and development and production. Summary of the Invention

[0006] The purpose of the present invention is to solve the defects such as damage, poisoning, and sub-healthy state of cells used for treatment, production, and research, and to provide high-quality healthy cells.

[0007] The technical solution of the present invention for achieving the above object is as follows: The cell damage repair incubator is composed of a support and fixation system A, a conditioning and culture system B, a filtration and circulation system C, and an oxygen supply and ventilation system D. Installation and disassembly can be conveniently carried out between and within different systems to facilitate cleaning and sterilization. The support and fixation system A includes a base A1, self-tapping screws A2, A3, a bracket A4, a bolt A5, and a nut A6; the conditioning and culture system B includes a culture tank B1, screws B2, a lid B3, a sealing ring B4, and a nut B5; the filtration and circulation system C includes a peristaltic pump C1, hoses C2, C3, a conical filter screen C4, a universal rotating shaft C5, a suction and discharge pipe C6, a conical filter screen C7, a universal rotating shaft C8, and a suction and discharge pipe C9; the oxygen supply and ventilation system D includes a sterilizing filter D1, a connecting hose D2, and a ventilation pipe D3.

[0008] Hoses C2 and C3 are the two ends of the same hose. The suction and discharge pipes C6 and C9 can both suck and discharge the cell culture medium. The conical filter screens C4 and C7 are conical mesh cylinders with open tops, and the mesh hole diameter is <5 μm to ensure that cultured cells do not enter the hose when the filtration and circulation system C is working. To prevent the cultured cells from clogging the mesh holes, when the filtration and circulation system C is working, the universal rotating shafts C5 and C8 can respectively ensure that the conical filter screens C4 and C7 rotate freely in the culture medium.

[0009] Except for the entire support and fixation system A and the peristaltic pump C1 in the filtration and circulation system C, the rest can be sterilized by high-pressure steam (103.4 kPa, 121.3 °C, 15 - 20 min). The culture tank B1 is made of high-temperature and high-pressure resistant borosilicate glass, which is transparent and convenient for observing the cell culture situation; the support and fixation system A, as well as the suction and discharge pipes C6, C9, conical filter screens C4, C7, universal rotating shafts C5, and C8 in the filtration and circulation system C, are all made of medical-grade stainless steel.

[0010] The advantages of the technical solution of the present invention include: ① The cell damage repair incubator uses its own culture solution to circulate and stir the culture, with gentle and uniform stirring, causing no damage to the cultured cells; ② The design of the cell damage repair incubator avoids installing probes for pH, dissolved oxygen, temperature, etc., reducing the chance of contamination, avoiding the adhesion of cultured cells to the probe part immersed in the culture solution, simplifying the operation, and reducing costs; ③ The cell damage repair incubator does not have its own heating system and is placed in a carbon dioxide incubator or a constant temperature incubator during operation. By adjusting the temperature of the carbon dioxide incubator or the constant temperature incubator, the optimal temperature required for culturing cells can be met, with gentle temperature changes and no damage to cells; ④ The universal rotating shaft design of the cell damage repair incubator helps prevent the cultured cells from clogging the conical filter screen; ⑤ The spherical design at the bottom of the culture tank of the cell damage repair incubator has good hydrodynamic advantages, avoiding the retention of cultured cells at the bottom; ⑥ The cell damage repair incubator is convenient to operate, inexpensive, and easy to promote and apply. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Attached Figure 1 is a perspective view of the present invention.

[0012] In the figure, A, support and fixation system, A1, base, A2, self-tapping screw, A3, self-tapping screw, A4, bracket, A5, bolt, A6, nut; B, conditioning and culturing system, B1, culture tank, B2, screw, B3, lid, B4, sealing ring, B5, nut; C, filtration and circulation system, C1, peristaltic pump, C2, hose, C3, hose, C4, conical filter screen, C5, universal rotating shaft, C6, suction and discharge pipe, C7, conical filter screen, C8, universal rotating shaft, C9, suction and discharge pipe; D, oxygen supply and ventilation system, D1, sterilizing filter, D2, connecting hose, D3, ventilation pipe. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0013] Embodiment 1:

[0014] Installation and use of the present invention.

[0015] As shown in the figure, the cell damage repair incubator is composed of a support and fixation system A, a conditioning and culturing system B, a filtration and circulation system C, and an oxygen supply and ventilation system D. The support and fixation system A includes a base A1, self-tapping screws A2, A3, a bracket A4, a bolt A5, and a nut A6; the conditioning and culturing system B includes a culture tank B1, screws B2, a lid B3, a sealing ring B4, and a nut B5; the filtration and circulation system C includes a peristaltic pump C1, hoses C2, C3, conical filter screens C4, C7, universal rotating shafts C5, C8, and suction and discharge pipes C6, C9; the oxygen supply and ventilation system D includes a sterilizing filter D1, a connecting hose D2, and a ventilation pipe D3.

[0016] Before use, first clean the culture tank B1, conical filter C4, universal rotating shaft C5, suction and discharge pipe C6, conical filter C7, universal rotating shaft C8 and suction and discharge pipe C9 according to the requirements of culturing cells, and then siliconize them with 3% methyl silicone resin alcohol solution to prevent cells from adhering to them during cell culture. Then, sterilize each part of the cell damage repair incubator by high-pressure steam (103.4 kPa, 121.3 °C, 15 - 20 min), except for the support and fixation system A and the peristaltic pump C1 in the filtration and circulation system C.

[0017] Installation and use steps: (1) In a laminar flow hood or biological safety cabinet, perform aseptic operation: ① Connect the pre-sterilized conical filter C4, universal rotating shaft C5, and suction and discharge pipe C6, install the suction and discharge pipe C6 on the lid B3, and connect the other end to the hose C3; ② Connect the pre-sterilized conical filter C7, universal rotating shaft C8, and suction and discharge pipe C9, install the suction and discharge pipe C9 on the lid B3, and connect the other end to the hose C2; ③ Connect the sterilizing filter D1, connecting hose D2, and ventilation pipe D3, and install the ventilation pipe D3 on the lid B3; ④ Install the sealing ring B4 on the lid B3; ⑤ Add the cell sample that needs to be repaired, detoxified, and restored to the culture tank B1, and then add the corresponding culture medium, reagents, and drugs according to the number of cells (the types and amounts added are determined according to specific needs, but the reagents and drugs added must be pre-filtered and sterilized); ⑥ Install the lid B3 on the culture tank B1 and tighten the screws appropriately. (2) Fix the bracket A4 to the base A1 with self-tapping screws A2 and A3; (3) Place the assembled culture tank B1 inside the circle of the bracket A4, manually install the bolt A5 and nut A6, and tighten appropriately; (4) Load the hoses C2 and C3 into the peristaltic pump C1 and place it on the base A1; (5) Place the installed cell damage repair incubator in a carbon dioxide incubator or a constant temperature incubator, adjust the temperature to the optimal temperature for cell growth (37 °C for human and mammalian cells), and adjust the rotation speed of the peristaltic pump to make the cell culture mixture move gently; (6) The cell conditioning culture time is usually several hours to dozens of hours. If the culture medium needs to be changed in the middle, take the cell damage repair incubator out of the incubator, and in the laminar flow hood or biological safety cabinet, through aseptic operation, use the peristaltic pump to draw out an appropriate amount (usually 50%) of the consumed culture medium from the suction and discharge pipe C6 or suction and discharge pipe C9, and then add an equal amount of fresh culture medium. If other reagents or drugs need to be added, after filtration and sterilization, add them together with the fresh culture medium; (7) Restore the original connection of the cell damage repair incubator, put it back into the incubator, adjust the temperature and the rotation speed of the peristaltic pump, and continue the culture.

[0018] The above technical solutions only reflect the preferred technical solutions of the present invention. Some changes that those skilled in the art may make to some parts thereof all reflect the principles of the present invention and fall within the protection scope of the present invention.

[0019] Example 2:

[0020] Repair of human amniotic mesenchymal stem cells after trypsin digestion injury.

[0021] For human amniotic mesenchymal stem cells cultured in a square flask, the cells basically cover the bottom of the flask. In a laminar flow hood or biosafety cabinet, perform aseptic operation, pour out the culture medium, add an appropriate amount of 0.25% trypsin solution, and place it in a 37°C constant temperature incubator for digestion. When there are signs of detachment of the cell layer at the bottom of the flask, pour out the trypsin solution and continue digestion using the residual solution. After the cell layer detaches, in a laminar flow hood or biosafety cabinet, perform aseptic operation, aspirate an appropriate amount of fresh culture medium, and repeatedly pipette the cells at the bottom of the flask to disperse them as much as possible to prepare a cell sample. Due to trypsin digestion and repeated pipetting, the sample cells will be damaged to a certain extent.

[0022] Repair steps: (1) In a laminar flow hood or biosafety cabinet, perform aseptic operation: ① Connect the pre-sterilized conical filter C4, universal joint C5, and suction and discharge tube C6, install the suction and discharge tube C6 on the lid B3, and connect the other end to the hose C3; ② Connect the pre-sterilized conical filter C7, universal joint C8, and suction and discharge tube C9, install the suction and discharge tube C9 on the lid B3, and connect the other end to the hose C2; ③ Connect the sterilizing filter D1, connecting hose D2, and ventilation tube D3, and install the ventilation tube D3 on the lid B3; ④ Install the sealing ring B4 on the lid B3; ⑤ Add the human amniotic mesenchymal stem cell sample to the culture tank B1, and then add the corresponding amount of serum-free medium, glutamine, and glucose according to the cell number; ⑥ Install the lid B3 on the culture tank B1 and tighten the screws appropriately. (2) Fix the bracket A4 to the base A1 with self-tapping screws A2 and A3; (3) Place the assembled culture tank B1 inside the circle of the bracket A4, manually install the bolt A5 and nut A6, and tighten appropriately; (4) Insert the hoses C2 and C3 into the peristaltic pump C1 and place it on the base A1; (5) Place the installed cell injury repair incubator in a carbon dioxide incubator or a constant temperature incubator, adjust the temperature to 37°C, and adjust the rotation speed of the peristaltic pump to make the cell culture mixture flow gently; (6) After 1 - 2 days of constant temperature culture, the injury repair is completed. Compared with before the repair culture, the cell morphology and proliferation ability are significantly enhanced, making it suitable for direct transplantation treatment.

Claims

1. A cell damage repair incubator, characterized in that, the cell damage repair incubator is composed of a support and fixation system (A), a conditioning and culture system (B), a filtration and circulation system (C), and an oxygen supply and ventilation system (D); the support and fixation system (A) includes a base (A1), self-tapping screw 1 (A2), self-tapping screw 2 (A3), a bracket (A4), a bolt (A5), and nut 1 (A6); the conditioning and culture system (B) includes a culture tank (B1), screws (B2), a lid (B3), a sealing ring (B4), and nut 2 (B5); the filtration and circulation system (C) includes a peristaltic pump (C1), hose 1 (C2), hose 2 (C3), conical filter 1 (C4), universal rotating shaft 1 (C5), suction and discharge pipe 1 (C6), conical filter 2 (C7), universal rotating shaft 2 (C8), and suction and discharge pipe 2 (C9); the oxygen supply and ventilation system (D) includes a sterilizing filter (D1), a connecting hose (D2), and a ventilation pipe (D3): conical filter 1 (C4) and conical filter 2 (C7) are conical mesh cylinders with open tops, and the mesh hole diameter is <5 μm to ensure that cultured cells do not enter the hose when the filtration and circulation system (C) is working.

2. The cell damage repair incubator according to claim 1, characterized in that, both ends of hose 1 (C2) and hose 2 (C3) are of the same hose.

3. The cell damage repair incubator according to claim 1, characterized in that, suction and discharge pipe 1 (C6) and suction and discharge pipe 2 (C9) can both suck and discharge cell culture medium.

4. The cell damage repair incubator according to claim 1, characterized in that, in order to prevent the cultured cells from clogging the mesh holes, when the filtration and circulation system (C) is working, universal rotating shaft 1 (C5) and universal rotating shaft 2 (C8) can respectively ensure that conical filter 1 (C4) and conical filter 2 (C7) rotate freely in the culture medium.

5. The cell damage repair incubator according to claim 1, characterized in that, except for the entire support and fixation system (A) and the peristaltic pump (C1) in the filtration and circulation system (C), the rest can be sterilized by high-pressure steam, and the sterilization parameters are 103.4 kPa, 121.3 °C, and 15 - 20 min.

6. The cell damage repair incubator according to claim 1, characterized in that, the culture tank (B1) is made of high-temperature and high-pressure resistant borosilicate glass, transparent, and is convenient for observing the cell culture situation.

Images