Two-phase circulating cell culture device

Abstract

The utility model relates to the technical field of cell culture, and particularly discloses a two-phase circulating cell culture device which comprises a culture cylinder, a liquid collecting box, macroporous resin and a circulating assembly, the culture cylinder comprises an outer cylinder, a second opening is formed in the top of the outer cylinder, an inner cylinder is sleeved with the outer cylinder, a first opening is formed in the top of the inner cylinder, a gap is formed between the inner cylinder and the outer cylinder, macroporous resin is placed in the gap, a cell secretion through hole is formed in the side wall of the inner cylinder, and a mixed liquid through hole is formed in the bottom of the outer cylinder; the liquid collecting box is arranged below the culture cylinder, the liquid collecting box comprises an outer cylinder liquid collecting box, the top of the outer cylinder liquid collecting box is provided with a mixed liquid input port, the mixed liquid input port is communicated with the mixed liquid through hole, and the side wall of the outer cylinder liquid collecting box is provided with a mixed liquid output port; the circulating assembly comprises a circulating pump and a circulating pipeline, the input end of the circulating pipeline is communicated with the mixed liquid output port, the output end of the circulating pipeline is communicated with the first opening, and the circulating pump is used for conveying the mixed liquid in the liquid collecting box of the outer cylinder to the inner cylinder.

Description

Technical Field

[0001] The utility model relates to the technical field of cell culture, in particular to a two-phase circulation cell culture device. Background Art

[0002] Paclitaxel, also known as taxol, taxotere, purple element, special element, is the most excellent natural anti-cancer drug discovered so far and has been widely used in the clinical treatment of breast cancer, ovarian cancer, some head and neck cancers and lung cancers. Paclitaxel is a natural secondary metabolite isolated and purified from the bark of the gymnosperm Taxus chinensis. In recent years, the global population and the incidence of cancer have increased explosively, and the demand for paclitaxel has also increased significantly. The paclitaxel required for clinical and scientific research is mainly extracted directly from Taxus chinensis. Since the content of paclitaxel in the plant body is quite low, about 1g of paclitaxel can be extracted from 13.6kg of bark. Treating a patient with ovarian cancer requires 3-12 Taxus chinensis trees over a hundred years old, which has led to a large amount of deforestation of Taxus chinensis, making this precious tree species on the verge of extinction. In addition, the resources of Taxus itself are very scarce, and the plants of the genus Taxus grow slowly, which has caused great difficulties for the further development and utilization of paclitaxel.

[0003] At present, the domestic market mainly uses organic extraction method and ultrasonic extraction method to extract paclitaxel. Among them, the organic extraction method uses the most common ethanol extraction. Under the current best conditions for paclitaxel extraction, the paclitaxel content only reaches 0.0125mg / g. However, in the extraction process of both methods, chemical pollution will be generated, and there are also problems such as long time consumption, high energy consumption and low extraction rate. In addition, the slow growth and poor regeneration ability of Taxus chinensis itself have greatly restricted the extraction of paclitaxel from plant tissues. And now many studies on the process of extracting paclitaxel focus more on the downstream extraction and synthesis, without paying attention to promoting the yield of paclitaxel secreted by Taxus chinensis cells in the upstream. When promoting the secretion of paclitaxel by Taxus chinensis cells in the upstream, the Taxus chinensis cell suspension is often discarded after only one secretion, resulting in a low extraction rate of paclitaxel. Content of the Utility Model

[0004] In order to solve the problem of low extraction rate of paclitaxel in the prior art, the utility model provides a two-phase circulation cell culture device.

[0005] The technical solution adopted by the present utility model is as follows: A two-phase circulating cell culture device, comprising: a culture tank, a liquid collection tank, macroporous resin, and a circulation assembly; the culture tank includes an outer tank, the top of the outer tank is provided with a second opening, an inner tank is sleeved inside the outer tank, the top of the inner tank is provided with a first opening, there is a gap between the inner tank and the outer tank, the macroporous resin is placed in the gap, the side wall of the inner tank is provided with cell secretion through holes, and the bottom of the outer tank is provided with mixed liquid through holes; the liquid collection tank is arranged below the culture tank, the liquid collection tank includes an outer tank liquid collection tank, the top of the outer tank liquid collection tank is provided with a mixed liquid input port, the mixed liquid input port is communicated with the mixed liquid through hole, and the side wall of the outer tank liquid collection tank is provided with a mixed liquid output port; the circulation assembly includes a circulation pump and a circulation pipeline, the input end of the circulation pipeline is communicated with the mixed liquid output port, the output end of the circulation pipeline is communicated with the first opening, and the circulation pump is used to transport the mixed liquid in the outer tank liquid collection tank to the inner tank.

[0006] Further, the side wall of the inner tank and the bottom wall of the outer tank are both made of cellulose acetate material, and the mixed liquid through holes and the cell secretion through holes are pores formed by cellulose acetate material.

[0007] Further, the tops of the inner tank and the outer tank are flush, a cover plate is arranged above the outer tank, and the cover plate is used to cover the inner tank and the outer tank; an eluent input hole corresponding to the second opening is opened on the cover plate; a cell liquid input hole and a mixed liquid input hole corresponding to the first opening are opened on the cover plate; the cell liquid input hole, the mixed liquid input hole, and the eluent input hole all penetrate through the cover plate.

[0008] Further, it further includes a stirring mechanism, a gas circulation assembly, and a heat preservation mechanism; wherein, the stirring mechanism includes a driving member and a stirring rod, the output end of the driving member is connected to the stirring rod, and the stirring rod extends vertically downward from above the inner tank into the inner tank; the gas circulation assembly includes an air distributor and an exhaust valve, the air distributor is arranged at the bottom of the inner tank, and the air distributor is connected to a gas source, and the exhaust valve is arranged on the corresponding cover plate of the inner tank; the heat preservation mechanism includes a heat preservation shell and a heating wire, and the heating wire is located between the outer wall of the outer tank and the inner wall of the heat preservation shell; a temperature sensor is arranged inside the inner tank, a temperature controller is arranged on the outer wall of the heat preservation shell, the temperature controller is connected to the temperature sensor, and the temperature sensor is used to detect the temperature of the cell suspension inside the inner tank.

[0009] Further, the distance between the stirring end of the stirring rod and the bottom of the inner tank is between 1 / 3 and 1 / 2 of the height of the inner tank.

[0010] Further, a number of tissue culture lamps are arranged on the side of the stirring rod facing the inner tank, and the tissue culture lamps are used to irradiate the cell suspension inside the inner tank.

[0011] Further, a first output port is provided at the bottom of the inner cylinder; the liquid collection tank further includes an inner cylinder liquid collection tank which is located below the inner cylinder. A cell liquid input port corresponding to the first output port is provided at the top of the inner cylinder liquid collection tank, and the inner cylinder liquid collection tank is sleeved inside the outer cylinder liquid collection tank.

[0012] Further, a solenoid valve is provided at the first output port, and a liquid discharge port is provided at the bottom of the inner cylinder liquid collection tank. The liquid discharge port is used for discharging the cell suspension.

[0013] The beneficial effects of the present utility model are as follows: By dividing the culture cylinder into an inner cylinder and an outer cylinder, and placing macroporous resin for adsorbing secretions in the outer cylinder, the cell secretions in the inner cylinder can be directly sucked out, without the need to discharge the cell suspension and then adsorb it through the macroporous resin, thus simplifying the adsorption process; By providing an outer cylinder liquid collection tank at the discharge port below the outer cylinder, the eluent and secretions are discharged into the outer cylinder liquid collection tank, and then through the setting of the circulation component, the eluent and secretions are re-transported back to the inner cylinder for continued culture and secretion in the inner cylinder. Compared with the prior art where the cell suspension is discarded after only one secretion, the cells in the cell suspension in this application can be subjected to multiple cycles of culture and secretion, thereby obtaining more cell secretions and improving the extraction rate. Description of the Drawings

[0014] Figure 1 is the overall schematic diagram of a two-phase circulation cell culture device of the present utility model;

[0015] Figure 2 is Figure 1 the perspective view of

[0016] Figure 3 the structural schematic diagram of the cylinder body;

[0017] Figure 4 the structural schematic diagram of the liquid collection tank;

[0018] Figure 5 the structural schematic diagram of the cover plate;

[0019] Figure 6 the structural schematic diagram of the heat preservation component.

[0020] Reference numerals: 1, culture tank; 101, inner tank; 102, outer tank; 103, first opening; 104, cell secretion through-hole; 105, second opening; 106, mixed liquid through-hole; 107, cover plate; 108, cell liquid input hole; 109, mixed liquid input hole; 110, eluent input hole; 111, first output port; 112, air hole; 2, liquid collection tank; 201, inner tank liquid collection tank; 202, outer tank liquid collection tank; 203, mixed liquid input port; 204, mixed liquid output port; 205, cell liquid input port; 3, macroporous resin; 4, circulation assembly; 401, circulation pump; 402, circulation pipeline; 5, stirring mechanism; 501, driving member; 502, stirring rod; 503, tissue culture lamp; 601, air distributor; 602, exhaust valve; 603, air pipe; 7, heat preservation mechanism; 701, heat preservation shell; 702, heating wire; 703, heat preservation material; 8, temperature sensor; 9, temperature controller; 10, drain port. Detailed implementation manners

[0021] In order to make the objectives, technical solutions and advantages of the present utility model clearer and more understandable, the present utility model will be further described in detail below with reference to the embodiments and the drawings. The illustrative embodiments and descriptions thereof of the present utility model are only used to explain the present utility model and do not limit the present utility model.

[0022] In the following description, a large number of specific details are set forth in order to provide a thorough understanding of the present utility model. However, it is obvious to those of ordinary skill in the art that the present utility model does not have to employ these specific details. In other instances, well-known structures, circuits, materials or methods have not been specifically described in order to avoid obscuring the present utility model.

[0023] Throughout the specification, references to "one embodiment", "embodiment", "one example" or "example" mean that the specific features, structures or characteristics described in connection with the embodiment or example are included in at least one embodiment of the present utility model. Thus, the phrases "one embodiment", "embodiment", "one example" or "example" appearing throughout the specification do not necessarily all refer to the same embodiment or example. Additionally, the specific features, structures or characteristics may be combined in any suitable combination and / or sub-combination in one or more embodiments or examples. Moreover, those of ordinary skill in the art should understand that the diagrams provided herein are for illustrative purposes only and are not necessarily drawn to scale. The term "and / or" used herein includes any and all combinations of one or more of the associated listed items.

[0024] In the description of the present utility model, it should be understood that the orientation or positional relationships indicated by the terms "front", "rear", "left", "right", "up", "down", "vertical", "horizontal", "high", "low", "inner", "outer", etc. are based on the orientation or positional relationships shown in the drawings. These are only for the convenience of describing the present utility model and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, it should not be construed as limiting the protection scope of the present utility model.

[0025] The following will Figures 1-6 describe the embodiments of the present utility model in detail with reference to the accompanying drawings.

[0026] Embodiment 1

[0027] At present, organic extraction method and ultrasonic extraction method are mainly used in the domestic market to extract paclitaxel. Among them, the organic extraction method uses the most common ethanol extraction. Under the current best conditions for paclitaxel extraction, the paclitaxel content only reaches 0.0125 mg / g. However, in the extraction process of both methods, chemical pollution will be generated, and there are problems such as long time consumption, high energy consumption, and low extraction rate. In addition, the slow growth and poor regeneration ability of Taxus plants themselves greatly limit the extraction of paclitaxel from plant tissues. And now many studies on the process of extracting paclitaxel focus more on the downstream extraction and synthesis, rather than paying attention to promoting the yield of paclitaxel secreted by Taxus cells in the upstream. When promoting the secretion of paclitaxel by Taxus cells in the upstream, the Taxus cell suspension is often discarded after only one secretion, resulting in a low extraction rate of paclitaxel.

[0028] To solve the above problems, this embodiment provides a two-phase circulation cell culture device. As Figures 1-4 shown, it includes: a culture tank 1, a liquid collection tank 2, a macroporous resin 3, and a circulation component 4; the culture tank 1 includes an outer tank 102, a second opening 105 is provided at the top of the outer tank 102, an inner tank 101 is sleeved inside the outer tank 102, a first opening 103 is provided at the top of the inner tank 101, there is a gap between the inner tank 101 and the outer tank 102, the macroporous resin 3 is placed in the gap, a cell secretion through hole 104 is provided on the side wall of the inner tank 101, and a mixed liquid through hole 106 is provided at the bottom of the outer tank 102;

[0029] It can be understood that the culture tank 1 is a double-layer structure, the outer tank 102 and the inner tank 101 share the middle layer, and the culture tank 1 is an open cylinder. The open mouth of the inner tank 101 is the first opening 103, and the open mouth of the outer tank 102 is the second opening 105; when cell culture liquid needs to be cultured, the cell suspension is directly input into the inner tank 101 from the first opening 103, and the eluent is directly input into the outer tank from the second opening 105.

[0030] In order to achieve the circulating culture of cell sap, a liquid collection tank 2 is arranged below the culture tank 1. Similarly, the liquid collection tank 2 includes an outer cylinder liquid collection tank 202. A mixed liquid input port 203 is provided at the top of the outer cylinder liquid collection tank 202. The mixed liquid input port 203 is communicated with a mixed liquid through hole 106. A mixed liquid output port 204 is provided on the side wall of the outer cylinder liquid collection tank 202. The macroporous resin 3 is arranged in the outer cylinder 102 and is used for adsorbing cell secretions.

[0031] Here, the shape of the box body of the liquid collection tank is not limited. The culture tank and the liquid collection tank can be connected and fixed by means of plugging, snap connection, screw connection or even welding. The culture tank and the liquid collection tank can be two separate parts as long as the liquid collection tank is located below the culture tank. Since the eluent and cell secretions in the outer cylinder need to return to the inner cylinder after elution, the mixed liquid through hole 106 at the bottom of the outer cylinder 102 is communicated with the mixed liquid input port 203.

[0032] The circulation component 4 includes a circulation pump 401 and a circulation pipeline 402. The input end of the circulation pipeline 402 is communicated with the mixed liquid output port 204. The output end of the circulation pipeline 402 is communicated with the first opening 103. The circulation pump 401 is used for conveying the materials in the outer cylinder liquid collection tank 202 to the inner cylinder 101.

[0033] The circulation process is as follows: The cell suspension is first cultured in the inner cylinder 101, then the cell secretions are adsorbed by the macroporous resin 3 in the outer cylinder 102, then the macroporous resin 3 is eluted by the eluent, and then it flows back to the inner cylinder 101 through the outer cylinder liquid collection tank 202 and the circulation pipeline 402. Such circulation realizes the circulating culture and secretion of the cell suspension and improves the yield of the obtained cell secretions. Here, the circulation pipeline 402 can be arranged on the outer wall of the outer cylinder 102 or can be arranged separately from the outer cylinder 102.

[0034] When using this two-phase circulating cell culture device, taking the Taxus cell suspension as an example, first place the Taxus cell suspension in the inner cylinder 101, and at the same time place the macroporous resin 3 in the outer cylinder 102. As the culture progresses, the Taxus cells will secrete paclitaxel, and the secreted paclitaxel is adsorbed by the macroporous resin 3 through the cell secretion through hole 104. After a period of time, inject the eluent ethanol into the outer cylinder 102 to elute the paclitaxel adsorbed on the macroporous resin 3. The eluent after elution and the eluted paclitaxel flow into the outer cylinder liquid collection tank 2 in the outer cylinder 102 through the mixed liquid through hole 106 at the bottom of the outer cylinder 102, and then start the circulation pump 401 to suck the eluent and the eluted paclitaxel back into the inner cylinder 101.

[0035] It can be understood that in this embodiment, paclitaxel and ethanol are made to flow back from the outer cylinder 102 to the inner cylinder 101, and then the above-mentioned culture and elution steps are repeated to achieve the cyclic culture of the Taxus cell suspension and obtain more paclitaxel. Compared with the existing method of discarding the cell culture solution after only one elution, more paclitaxel is obtained in this embodiment.

[0036] Example 2

[0037] In order to enable the paclitaxel in the inner cylinder 101 to be adsorbed by the macroporous resin 3 in the outer cylinder 102 while the Taxus cells in the inner cylinder 101 do not enter the outer cylinder 102, in this embodiment, the side wall of the inner cylinder 101 and the bottom wall of the outer cylinder 102 are both made of cellulose acetate. That is, the intermediate layer between the inner cylinder and the outer cylinder is made of cellulose acetate, and the mixed liquid through holes 106 and the cell secretion through holes 104 are pores formed by cellulose acetate. In this way, the cell secretion paclitaxel can be directly adsorbed by the macroporous resin 3 in the outer cylinder 102 through the intermediate layer. During elution, the eluent, paclitaxel, and cell liquid (substances other than Taxus cells) flow into the outer cylinder liquid collection box 202 through the mixed liquid through holes 106 at the bottom of the outer cylinder 102.

[0038] Example 3

[0039] In order to prevent bacteria in the air from entering the inner cylinder 101 and the outer cylinder 102 during cell culture and contaminating the Taxus cell suspension and paclitaxel, the tops of the inner cylinder 101 and the outer cylinder 102 are flush, and a cover plate 107 is provided above the outer cylinder 102. The cover plate 107 is used to cover the inner cylinder 101 and the outer cylinder 102; an eluent input hole 110 corresponding to the second opening 105 is opened on the cover plate 107; a cell liquid input hole 108 and a mixed liquid input hole 109 corresponding to the first opening 103 are opened on the cover plate 107; the cell liquid input hole 108, the mixed liquid input hole 109, and the eluent input hole 110 all penetrate through the cover plate 107. As Figure 2 and 5 shown, the culture cylinder 1 further includes a cover plate 107. The cover plate 107 is provided with a cell liquid input hole 108, a mixed liquid input hole 109, and an eluent input hole 110 penetrating therethrough; wherein, the cell liquid input hole 108 and the mixed liquid input hole 109 are both communicated with the first opening 103, the eluent input hole 110 is communicated with the second opening 105, and the cover plate 107 covers the tops of the inner cylinder 101 and the outer cylinder 102.

[0040] For reference, on the one hand, the cover plate 107 and the culture cylinder 1 are two relatively separate components. The cover plate 107 can be fixedly connected to the top of the outer cylinder 102 by means of hinge, so as to obtain better maintenance convenience. On the other hand, the cover plate 107 and the culture cylinder 1 adopt an integrated structure. By fixing the cover plate 107 to the top of the outer cylinder 102, the cover plate 107 and the culture cylinder 1 have a high degree of consistency, so as to ensure the relative position relationship of the cell liquid input hole 108, the mixed liquid input hole 109 and the eluent input hole 110, and ensure the relative stability of the cover plate 107 with respect to the first opening 103 and the second opening 105. The above two specific installation methods of the cover plate 107 are for adaptive selection without limitation. In principle, as long as the function of culturing cells is satisfied.

[0041] Example 4

[0042] In order to increase the yield of paclitaxel, as Figure 2 shown, the cell culture device further includes a stirring mechanism 5, a gas circulation assembly and a heat preservation mechanism 7; the stirring mechanism 5 includes a driving member 501 and a stirring rod 502, the output end of the driving member 501 is connected to the stirring rod 502, and the stirring rod 502 extends vertically downward from above the inner cylinder 101 into the inner cylinder 101;

[0043] For reference, the driving member 501 can be any device that can provide rotational power for the stirring rod 502, such as a motor. It can be understood that: the upper end of the stirring rod 502 is the fixed end, the lower end of the stirring rod 502 is the stirring end, the fixed end is fixedly connected to the output shaft of the driving member 501, the driving member 501 drives the stirring rod 502 to rotate, the stirring end extends into the inner cylinder 101 through the cell liquid input hole 108, and the outer diameter of the fixed end is equal to the inner diameter of the cell liquid input hole 108, so as to prevent bacteria in the air from entering the inner cylinder 101.

[0044] In addition, since too slow stirring speed may lead to uneven distribution of nutrients and oxygen in the culture solution, affecting cell growth; too fast stirring speed will generate too large shear force, causing damage to cells. Therefore, when stirring the Taxus cell suspension, the rotation speed of the stirring rod 502 is set to 50 - 70 rpm, preferably 60 rpm.

[0045] The gas circulation assembly includes an air distributor 601 and an exhaust valve 602. The air distributor 601 is arranged at the bottom of the inner cylinder 101, and the air distributor 601 is connected to a gas source. The exhaust valve 602 is arranged on the cover plate 107 corresponding to the inner cylinder 101;

[0046] For reference, the air distributor 601 is a prior art, and its structure will not be described in detail. However, it should be noted that the air distributor 601 is connected to an external air source through an air pipe 603. Therefore, the air pipe 603 also needs to pass through the cover plate 107. That is to say, there is also an air hole 112 opened on the cover plate 107. And since gas is generated during the cell culture process, the exhaust valve 602 is used to discharge the gas in the inner cylinder 101. Similarly, the exhaust valve 602 is a prior art, and its structure will not be described in detail.

[0047] In order to further increase the yield of paclitaxel secreted by Taxus cells, as Figure 4 shown, the heat preservation mechanism 7 includes a heat preservation shell 701 and a heating wire 702. The heating wire 702 is located between the outer wall of the outer cylinder 102 and the inner wall of the heat preservation shell 701. A temperature sensor 8 is arranged in the inner cylinder 101, and a temperature controller 9 is arranged on the outer wall of the heat preservation shell 701. The temperature controller 9 is connected to the temperature sensor 8, and the temperature sensor 8 is used to detect the temperature of the cell suspension in the inner cylinder 101.

[0048] For reference, the heating wire 702 is attached to the outer wall of the outer cylinder 102, and the heat preservation shell 701 is attached to the heating wire 702 and wraps the outer cylinder 102. The heating wire 702 is controlled by the temperature controller 9, and the temperature controller 9 is connected to the temperature sensor 8 to monitor the temperature change in the Taxus cell suspension in real time. In order to facilitate the secretion of paclitaxel by Taxus cells, in this embodiment, the temperature of the heating wire 702 is controlled between 18°C and 25°C, preferably 20°C. In order to obtain a better heat preservation effect, a heat preservation material 703 such as asbestos, mineral wool, phenolic resin foaming material, etc. can also be arranged between the heating wire 702 and the heat preservation shell 701.

[0049] Through the above settings of the rotation speed and temperature of the stirring rod 502, the Taxus cell suspension can secrete paclitaxel to the greatest extent during the culture process, further increasing the yield and extraction rate of paclitaxel.

[0050] Example 5

[0051] Since the Taxus cells are mainly concentrated at the bottom of the inner cylinder 101 during culture, in order to ensure that the stirring rod 502 does not directly contact the cells during stirring, and at the same time can effectively stir the Taxus cell suspension and maintain the uniform distribution of nutrients and oxygen, the distance between the stirring end of the stirring rod 502 and the bottom of the inner cylinder 101 is between 1 / 3 and 1 / 2 of the height of the inner cylinder 101.

[0052] Example 6

[0053] In order to further increase the yield of paclitaxel secreted by Taxus cells, in this embodiment, as Figure 2As shown in the figure, on the side of the stirring rod 502 facing the inner cylinder 101, there are several tissue culture lamps 503, and the tissue culture lamps 503 are used to irradiate the Taxus cell suspension in the inner cylinder 101.

[0054] Embodiment 7

[0055] Since during the cultivation process of the Taxus cell suspension, quality control of the Taxus cells is required, such as monitoring cell viability, growth status, etc. If the quality control is qualified, the cultivation continues; if not, a new Taxus cell suspension is replaced. Therefore, in this embodiment, as Figure 3 shown, a first output port 111 is provided at the bottom of the inner cylinder 101; the liquid collection tank 2 further includes an inner cylinder liquid collection tank 201, the inner cylinder liquid collection tank 201 is located below the inner cylinder 101, and a cell liquid input port 205 corresponding to the first output port 111 is provided at the top of the inner cylinder liquid collection tank 201. The inner cylinder liquid collection tank 201 is used to collect the Taxus cell suspension in the inner cylinder 101, and the inner cylinder liquid collection tank 201 is sleeved inside the outer cylinder liquid collection tank 202.

[0056] For reference, the inner cylinder liquid collection tank 201 can be connected to the inner cylinder 101 by means of clamping, plugging, threaded connection, welding, etc. The inner cylinder liquid collection tank 201 and the outer cylinder liquid collection tank 202 can be two separate boxes, or can be a box with a double-layer structure like the culture cylinder 1, and all can implement the above technical solutions. And due to the different functions of the inner cylinder liquid collection tank 201 and the outer cylinder liquid collection tank 202 during the cyclic cultivation process, it is preferably a separate box. In this case, when it is necessary to drain the cell liquid in the inner cylinder 101 for quality control, the inner cylinder liquid collection tank 201 can be directly separated from the inner cylinder 101.

[0057] In order to better control the cell liquid in the inner cylinder 101 to flow into the inner cylinder liquid collection tank 201, a solenoid valve (not shown in the figure) is provided at the first output port 111. And in order to facilitate the drainage of the Taxus cell suspension in the inner cylinder liquid collection tank 201, a drainage port 10 is provided at the bottom of the inner cylinder liquid collection tank 201.

[0058] The above embodiments only represent the specific implementation manners of the present invention, and the description is relatively specific and detailed, but it cannot be understood as a limitation on the scope of the patent of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several deformations and improvements can still be made, and these all belong to the protection scope of the present invention.

Claims

1. A two-phase circulation cell culture device, characterized in that: include: A culture tank (1), a liquid collection tank (2), a macroporous resin (3) and a circulation component (4); The culture cylinder (1) comprises an outer cylinder (102), a second opening (105) is provided at the top of the outer cylinder (102), an inner cylinder (101) is provided inside the outer cylinder (102), a first opening (103) is provided at the top of the inner cylinder (101), a gap is provided between the inner cylinder (101) and the outer cylinder (102), a macroporous resin (3) is placed in the gap, a cell secretion through hole (104) is provided on the side wall of the inner cylinder (101), and a mixed liquid through hole (106) is provided at the bottom of the outer cylinder (102); The liquid collecting box (2) is arranged below the culture cylinder (1), and the liquid collecting box (2) comprises an outer cylinder liquid collecting box (202), a mixed liquid input port (203) is provided on the top of the outer cylinder liquid collecting box (202), the mixed liquid input port (203) is connected to the mixed liquid through hole (106), and a mixed liquid output port (204) is provided on the side wall of the outer cylinder liquid collecting box (202); The circulation component (4) comprises a circulation pump (401) and a circulation pipeline (402); the input end of the circulation pipeline (402) is connected to the mixed liquid output port (204); the output end of the circulation pipeline (402) is connected to the first opening (103); and the circulation pump (401) is used to transport the mixed liquid in the outer cylinder liquid collecting tank (202) to the inner cylinder (101).

2. The two-phase circulation cell culture device according to claim 1, characterized in that: The side wall of the inner cylinder (101) and the bottom wall of the outer cylinder (102) are both made of cellulose acetate, and the mixed liquid through hole (106) and the cell secretion through hole (104) are pores formed by the cellulose acetate material.

3. The two-phase circulation cell culture device according to claim 1, characterized in that: The tops of the inner cylinder (101) and the outer cylinder (102) are flush with each other, and a cover plate (107) is provided above the outer cylinder (102), and the cover plate (107) is used to cover the inner cylinder (101) and the outer cylinder (102); The cover plate (107) is provided with an eluent input hole (110) corresponding to the second opening (105); The cover plate (107) is provided with a cell solution input hole (108) and a mixed solution input hole (109) corresponding to the first opening (103); The cell solution input hole (108), the mixed solution input hole (109) and the eluent input hole (110) all penetrate the cover plate (107).

4. The two-phase circulation cell culture device according to claim 1, characterized in that: It also includes a stirring mechanism (5), a gas circulation component and a heat preservation mechanism (7); The stirring mechanism (5) comprises a driving member (501) and a stirring rod (502), the output end of the driving member (501) is connected to the stirring rod (502), and the stirring rod (502) extends from the cell fluid input hole (108) into the inner cylinder (101); The gas circulation component comprises an air distributor (601) and an exhaust valve (602); the air distributor (601) is arranged at the bottom of the inner cylinder (101), and the air distributor (601) is connected to an air source; the exhaust valve (602) is arranged on a cover plate (107) corresponding to the inner cylinder (101); The heat preservation mechanism (7) comprises a heat preservation shell (701) and an electric heating wire (702), wherein the electric heating wire (702) is located between the outer wall of the outer cylinder (102) and the inner wall of the heat preservation shell (701); a temperature sensor (8) is arranged in the inner cylinder (101), and a temperature controller (9) is arranged on the outer wall of the heat preservation shell (701), wherein the temperature controller (9) is connected to the temperature sensor (8), and the temperature sensor (8) is used to detect the temperature of the cell suspension in the inner cylinder (101).

5. The two-phase circulation cell culture device according to claim 4, characterized in that: The distance between the stirring end of the stirring rod (502) and the bottom of the inner cylinder (101) is between 1 / 3 and 1 / 2 of the height of the inner cylinder (101).

6. The two-phase circulation cell culture device according to claim 4, characterized in that: A plurality of tissue culture lamps (503) are arranged on the side of the stirring rod (502) facing the inner cylinder (101), and the tissue culture lamps (503) are used to irradiate the cell suspension in the inner cylinder (101).

7. The two-phase circulation cell culture device according to claim 1, characterized in that: The bottom of the inner cylinder (101) is provided with a first output port (111); the liquid collecting box (2) further comprises an inner cylinder liquid collecting box (201), the inner cylinder liquid collecting box (201) is located below the inner cylinder (101), the top of the inner cylinder liquid collecting box (201) is provided with a cell fluid input port (205) corresponding to the first output port (111), and the inner cylinder liquid collecting box (201) is sleeved in the outer cylinder liquid collecting box (202).

8. The two-phase circulation cell culture device according to claim 7, characterized in that: The first output port (111) is provided with a solenoid valve, and the bottom of the inner cylinder liquid collecting box (201) is provided with a liquid discharge port (10), and the liquid discharge port (10) is used to discharge the cell suspension.

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