CN112940934B

CN112940934B - In-vitro life support perfusion culture system and control method thereof

Info

Publication number: CN112940934B
Application number: CN202110124977.4A
Authority: CN
Inventors: 王玄; 陈睿
Original assignee: Shanghai Ruiyu Biotech Co Ltd
Current assignee: Shanghai Ruiyu Biotech Co Ltd
Priority date: 2021-01-29
Filing date: 2021-01-29
Publication date: 2023-11-07
Anticipated expiration: 2041-01-29
Also published as: CN112940934A

Abstract

The invention relates to the technical field of in-vitro life culture, and discloses an in-vitro life support perfusion culture system, which comprises the following components: a premix unit comprising a premix chamber, wherein a nutrient solution and a gas are added into the premix chamber, and the mixture is mixed in the premix chamber to form a culture solution; the culture unit comprises a culture cavity, wherein a first membrane component is arranged in the culture cavity and divides the culture cavity into a first culture cavity and a second culture cavity; the first culture cavity is communicated with the premixing cavity, the premixing cavity can convey culture solution to the first culture cavity, the culture solution in the first culture cavity can flow back into the premixing cavity, and the second culture cavity is used for containing culture and culturing the culture; or the first culture cavity is communicated with the premixing cavity, the premixing cavity can convey culture solution to the first culture cavity, the second culture cavity is communicated with the premixing cavity, the culture solution in the second culture cavity can flow back into the premixing cavity, and the first culture cavity or the second culture cavity is used for containing culture and culturing the culture.

Description

In-vitro life support perfusion culture system and control method thereof

Technical Field

The invention relates to the technical field of in-vitro life culture, in particular to an in-vitro life support perfusion culture system and a control method thereof.

Background

The organoid is used as a 3D cell tissue and has quite high accuracy in the aspect of evaluating the essential drug effect of an evaluation tool of tumor drugs. At present, the organoid is mainly cultured in a static state in a pore plate, an operator shears the obtained primary sample, then coats the primary sample in matrigel at low temperature, then adds a culture medium, puts the primary sample into an incubator for culture, and performs liquid exchange for 3 times per week and performs passage for 7-10 days. This culture mode has a number of problems: firstly, the organoids have slow growth speed and long culture period, and the culture chamber is required to be opened periodically for manual liquid exchange in the operation process, so that the pollution risk is high; when the culture sample is increased, the cost investment of manpower, equipment, space and the like can be greatly increased. Second, static culture in the culture plate can lead to insufficient exchange of the organoids with the culture medium, especially insufficient absorption of internal nutrients by 3D cell tissues, and eventually can limit the growth of the organoids, limiting the extent of organoids and simulating the similarity of human organs.

In order to solve the technical problems, the prior art adopts perfusion culture, and the culture solution in the culture plate flows out at any time and is discharged, but the perfusion culture mode has lower utilization rate of the culture solution, so that the culture cost is higher.

Therefore, an in vitro life support perfusion culture system and a control method thereof are needed to solve the above technical problems.

Disclosure of Invention

The invention aims to provide an in-vitro life-support perfusion culture system and a control method thereof, which realize perfusion culture, improve the utilization rate of culture solution and reduce the culture cost.

To achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, there is provided an in vitro life support perfusion culture system comprising:

a premix unit comprising a premix chamber, wherein a nutrient solution and a gas are added into the premix chamber, and the mixture is mixed in the premix chamber to form a culture solution;

the culture unit comprises a culture cavity, wherein a first membrane component is arranged in the culture cavity and divides the culture cavity into a first culture cavity and a second culture cavity, and the first membrane component is used for intercepting and/or penetrating partial components in culture solution;

the first culture cavity is communicated with the premixing cavity, the premixing cavity can convey culture solution to the first culture cavity, the culture solution in the first culture cavity can flow back into the premixing cavity, and the second culture cavity is used for containing culture and culturing the culture; or (b)

The first culture cavity is communicated with the premixing cavity, the premixing cavity can convey culture solution to the first culture cavity, the second culture cavity is communicated with the premixing cavity, culture solution in the second culture cavity can flow back into the premixing cavity, and the first culture cavity or the second culture cavity is used for containing culture and culturing the culture.

As an optimal technical scheme of the in-vitro life support perfusion culture system, the first culture cavity is provided with a liquid inlet, the first culture cavity or the second culture cavity is provided with a liquid outlet, and the liquid inlet and the liquid outlet are communicated with the premixing cavity.

As a preferable technical scheme of the in-vitro life support perfusion culture system, the system further comprises a collection unit, wherein the collection unit is communicated with the liquid outlet and is used for collecting the culture solution discharged from the culture chamber.

As an optimized technical scheme of the in-vitro life support perfusion culture system, the system further comprises a three-way valve, wherein three interfaces of the three-way valve are respectively communicated with the liquid draining port, the collecting unit and the premixing chamber through pipelines.

As a preferable technical proposal of the in vitro life support perfusion culture system, the system also comprises a power unit,

The power unit is arranged between the premixing chamber and the liquid inlet; and/or

The power unit is arranged between the three-way valve and the liquid outlet.

As a preferred embodiment of the in vitro life support perfusion culture system, the premix unit further comprises a fluid replacement chamber in communication with the premix chamber for delivering a nutrient fluid into the premix chamber, the fluid replacement chamber being capable of directionally delivering one or more components required for the culture into the premix chamber.

As a preferable technical scheme of the in-vitro life support perfusion culture system, the first culture cavity is provided with a liquid inlet which is communicated with the premixing cavity, and the first culture cavity or the second culture cavity is provided with a liquid outlet;

the in vitro life support perfusion culture system further comprises:

the exchange unit comprises an exchange chamber and a second membrane assembly, the exchange chamber comprises a first exchange chamber and a second exchange chamber, the second membrane assembly is arranged between the first exchange chamber and the second exchange chamber and is communicated with the first exchange chamber through the second membrane assembly, and the liquid outlet and the premixing unit are both communicated with the first exchange chamber;

The fluid replacement unit is communicated with the second exchange cavity and is used for conveying a supplementary nutrient fluid to the second exchange cavity, the fluid replacement unit can directionally convey one or more components required by the culture into the second exchange cavity, and the components in the supplementary nutrient fluid can permeate into the first exchange cavity through the second membrane component.

As a preferred technical scheme of the in-vitro life support perfusion culture system, the system further comprises a collecting unit, wherein the collecting unit is communicated with the second exchange cavity and is used for collecting the culture solution discharged by the second exchange cavity.

In a second aspect, there is provided an in vitro life support perfusion culture system comprising:

the culture unit comprises a culture cavity, wherein a first membrane component is arranged in the culture cavity, the culture cavity is divided into a first culture cavity and a second culture cavity by the first membrane component, the first culture cavity is communicated with the premixing cavity, the premixing cavity can convey culture solution to the first culture cavity, and the first culture cavity is used for containing culture and culturing the culture;

A fluid replacement unit in communication with the second culture chamber for providing a nutrient fluid to the second culture chamber, the fluid replacement unit being capable of directionally delivering one or more components required for the culture into the second exchange chamber, at least a portion of the components in the nutrient fluid in the second culture chamber being permeable to the first culture chamber through the first membrane module;

the culture solution in the first culture cavity and/or the second culture cavity can be conveyed into the premixing cavity.

As a preferable technical scheme of the in-vitro life support perfusion culture system, the system further comprises a collection unit, wherein at least the second culture cavity is communicated with the collection unit in the first culture cavity and the second culture cavity.

As a preferred technical scheme of the in-vitro life support perfusion culture system, a third membrane component is arranged in the first culture cavity, the third membrane component divides the first culture cavity into a first sub-culture cavity and a second sub-culture cavity, the third membrane component is used for intercepting and/or penetrating components in culture solution, the third membrane component can also be used for intercepting the culture, and the first sub-culture cavity is communicated with the second culture cavity through the first membrane component;

The premixing cavity is communicated with the first sub-culture cavity, the culture solution in the premixing cavity can be conveyed into the first sub-culture cavity, and the second sub-culture cavity is used for placing a culture and culturing the culture; or (b)

The premixing cavity is communicated with the second sub-culture cavity, the culture solution in the premixing cavity can be conveyed into the second sub-culture cavity, and the first sub-culture cavity is used for placing a culture and culturing the culture.

As a preferred technical scheme of the in-vitro life support perfusion culture system, a third membrane component is arranged in the first culture cavity, the third membrane component divides the first culture cavity into a first sub-culture cavity and a second sub-culture cavity, the third membrane component is used for intercepting and/or penetrating components in culture solution, and the third membrane component can also be used for intercepting the culture;

the first sub-culture cavity and the second sub-culture cavity are communicated with the second culture cavity through the first membrane component, the premixing cavity is communicated with the first sub-culture cavity, culture solution in the premixing cavity can be conveyed into the first sub-culture cavity, and the second sub-culture cavity is used for placing a culture and culturing the culture.

As a preferred embodiment of the in vitro life support perfusion culture system, the first sub-culture chamber and/or the second sub-culture chamber is capable of delivering the culture fluid to the pre-mix unit.

As a preferred technical scheme of the in-vitro life support perfusion culture system, when the premixing chamber is communicated with the second sub culture chamber, the first sub culture chamber is used for placing a culture and culturing the culture, and the culture solution in the first sub culture chamber can be conveyed into the second sub culture chamber;

when the premixing chamber is communicated with the first sub-culture chamber, the second sub-culture chamber is used for placing a culture and culturing the culture, and the culture solution in the second sub-culture chamber can be conveyed into the first sub-culture chamber.

As an optimized technical scheme of the in-vitro life support perfusion culture system, the premixing unit further comprises a pH detection piece, a dissolved oxygen detection piece and a signal detector, wherein the pH detection piece and the dissolved oxygen detection piece are arranged in the premixing chamber, and the signal detector can sense signals fed back by the pH detection piece and the dissolved oxygen detection piece so as to obtain the pH value and the dissolved oxygen amount of the culture solution in the premixing chamber.

As a preferable technical scheme of the in-vitro life support perfusion culture system, the pH detection piece is a pH electrode plate, and the dissolved oxygen detection piece is a dissolved oxygen electrode plate;

the pH electrode plate is arranged on the inner wall of the premixing chamber or immersed in the liquid of the premixing chamber, and the dissolved oxygen detector is arranged on the inner wall of the premixing chamber or immersed in the liquid of the premixing chamber.

As a preferred embodiment of the in vitro life support perfusion culture system, the premix unit further comprises a gas premix control unit in communication with the premix chamber for delivering gas into the premix chamber.

As a preferred technical solution of the in vitro life support perfusion culture system, at least one of the pre-mixing unit, the culture chamber and the collection unit is a disposable consumable.

As a preferable technical scheme of the in-vitro life support perfusion culture system, a plurality of culture chambers are provided, and the culture chambers are communicated with the premixing chamber.

As a preferred technical scheme of the in vitro life support perfusion culture system, the system further comprises:

the temperature control unit comprises a refrigeration assembly and a temperature control module, wherein the refrigeration assembly is electrically connected with the temperature control module, and the temperature control module is used for controlling the refrigeration assembly to cool the culture chamber to a first preset temperature, and the first preset temperature is the liquefaction temperature of the culture supporting structure;

The culture chamber is provided with a sampling port, and the sampling port is used for taking out the culture after the culture supporting structure is liquefied.

As a preferred technical scheme of the in-vitro life support perfusion culture system, the temperature control unit further comprises a heating component, the heating component is electrically connected with the temperature control module, and the temperature control module is used for controlling the heating component to heat the culture chamber and/or the premixing chamber.

As an in vitro life support perfusion culture system's preferred technical scheme, the subassembly that heats includes a plurality of pieces that heat, the refrigeration subassembly includes a plurality of refrigeration pieces, a plurality of heat the piece interval setting, a plurality of the refrigeration piece interval setting is provided with one between two adjacent the piece that heats the piece, is provided with one between two adjacent refrigeration pieces and heats the piece, the energy of heating the piece can be transmitted to the refrigeration piece.

As a preferable technical scheme of the in-vitro life support perfusion culture system, the temperature control unit further comprises a temperature detection piece, wherein the temperature detection piece is electrically connected with the temperature control module and is used for detecting the temperature of the culture solution in the culture cavity.

a microscopic observation module comprising a stage on which the culture chamber is placed, the stage being capable of placing at least one of the culture chambers, and an observation assembly for observing a culture within the culture chamber;

the microscopic observation module further comprises a frame, the objective table is arranged on the frame in a swinging mode, and the objective table can drive the culture chamber to swing synchronously.

As an in vitro life support perfusion culture system's preferred technical scheme, microscopic observation module still includes autosampler and advances a kind track, the autosampler track sets up in the frame, the autosampler sets up slidingly advance kind track is last, the autosampler is used for to cultivate the cavity application of sample.

the sterile control module comprises a sterile working chamber, a filtering assembly and a sterilizing assembly, wherein at least the culture chamber is arranged in the sterile working chamber, the filtering assembly is used for filtering gas introduced into the sterile working chamber, and the sterilizing assembly is used for sterilizing the sterile working chamber.

As a preferred technical solution of the in vitro life support perfusion culture system, the culture unit further comprises a mixing module for shaking the culture solution in the culture chamber.

As an optimal technical scheme of the in-vitro life support perfusion culture system, the number of the premixing chambers is multiple, the number of the gas premixing control unit is one, and the gas premixing control unit is simultaneously communicated with the plurality of premixing chambers and can control the gas concentration in the plurality of premixing chambers.

In a third aspect, a control method of an in vitro life support perfusion culture system is provided, which is directed to the in vitro life support perfusion culture system, and includes the following steps:

detecting the concentration of a desired component of the culture in the culture chamber and/or determining the growth of the culture, and controlling the rate at which the premix chamber delivers culture fluid to the culture chamber based on the detected concentration and/or the determined growth of the culture.

In a fourth aspect, a control method of an in vitro life support perfusion culture system is provided, which is directed to the in vitro life support perfusion culture system as described above, comprising the steps of:

detecting the concentration of components required by the culture in the culture solution in the first culture cavity and/or determining the growth condition of the culture, and controlling the speed of conveying the culture solution to the first culture cavity by the premixing cavity and the speed of conveying the nutrient solution to the second culture cavity by the fluid supplementing unit according to the detected concentration and/or the determined growth condition of the culture.

The invention has the beneficial effects that:

the culture solution can be conveyed to the first culture cavity by the premixing cavity, and the culture solution in the first culture cavity or the second culture cavity can flow back into the premixing cavity, so that perfusion culture of the culture is realized, cyclic utilization of the culture solution is realized, utilization rate of the culture solution is improved, and culture cost is reduced.

In addition, the culture solution in the premixing cavity can be conveyed into the first culture cavity, the nutrient solution can be conveyed into the second culture cavity by the fluid supplementing unit, the first culture cavity and/or the culture solution in the second culture cavity can be conveyed into the premixing cavity, and components in the nutrient solution in the second culture cavity permeate into the first culture cavity through the membrane component for absorption and utilization of the culture; the first membrane component can intercept the components required by the growth of the culture in the culture solution in the first culture cavity, waste generated by the metabolism of the culture in the culture solution can permeate into the second culture cavity through the membrane component, the culture solution in the first culture cavity and/or the second culture cavity is reversed to the premixing cavity, and then the culture solution is conveyed into the first culture cavity for continuous use, so that the recycling of the culture solution is realized, the utilization rate of the culture solution is improved, and the culture cost is reduced. The components in the nutrient solution in the second culture cavity permeate into the first culture cavity through the membrane component for absorption and utilization of the culture, and can be directionally cultured according to the consumption condition of the components in the culture solution to supplement the components, so that the use amount of the nutrient solution is reduced, and the culture cost is reduced.

Moreover, the gas premixing control unit can be used for introducing gas into the premixing cavity to realize the adjustment of the dissolved oxygen concentration of the culture solution; the temperature control unit can control the temperature of the premixing chamber and the culture chamber, the pH value of the culture solution is regulated by adding the amount of the nutrient solution and the alkali liquor, and the temperature, the pH value and the dissolved oxygen concentration of the culture solution can be controlled in the premixing chamber, so that the system is free from the constraint of a carbon dioxide incubator, and the scale of culture can be arbitrarily enlarged.

Drawings

FIG. 1 is a schematic diagram of an in vitro life support perfusion culture system according to an embodiment of the present invention;

fig. 2 is a schematic structural diagram of a temperature control unit according to a first embodiment of the present invention;

fig. 3 is a schematic structural diagram II of a temperature control unit according to a first embodiment of the present invention;

FIG. 4 is a schematic diagram of a heating assembly and a cooling assembly according to a first embodiment of the present invention;

FIG. 5 is a schematic diagram of a microscopic observation module according to a first embodiment of the present invention;

FIG. 6 is a schematic diagram of a first embodiment of a microscopic observation module according to the present invention;

FIG. 7 is a schematic diagram of a microscopic observation module according to an embodiment of the present invention;

fig. 8 is a schematic structural view of a sterility control module according to a first embodiment of the present invention;

FIG. 9 is a schematic diagram of a microscopic observation module according to a third embodiment of the present invention;

FIG. 10 is a schematic diagram of a microscopic observation module according to a third embodiment of the present invention;

FIG. 11 is a schematic diagram of an in vitro life support perfusion culture system according to a fourth embodiment of the present invention;

FIG. 12 is a schematic diagram of an in vitro life support perfusion culture system according to a fifth embodiment of the present invention;

FIG. 13 is a schematic diagram of an in vitro life support perfusion culture system according to a sixth embodiment of the present invention;

FIG. 14 is a schematic diagram of an in vitro life support perfusion culture system according to a seventh embodiment of the present invention;

FIG. 15 is a schematic diagram of an in vitro life support perfusion culture system according to an eighth embodiment of the present invention;

FIG. 16 is a schematic view showing the structure of a culture chamber according to an eighth embodiment of the present invention;

FIG. 17 is a schematic diagram II of an in vitro life support perfusion culture system according to an eighth embodiment of the present invention;

FIG. 18 is a schematic diagram of an in vitro life support perfusion culture system according to a ninth embodiment of the present invention;

fig. 19 is a schematic structural diagram of an in vitro life support perfusion culture system according to a tenth embodiment of the present invention.

In the figure: 1. a premix unit; 101. a premix chamber; 102. a pH detecting member; 103. a dissolved oxygen detector; 104. a fluid replacement chamber; 105. a gas premix control unit;

2. A culture chamber; 201. a sample adding port; 202. a sampling port; 203. a first membrane module; 204. a three-way valve; 205. a first culture chamber; 2051. a first subculture chamber; 2052. a second subculture chamber; 206. a second culture chamber; 207. a third membrane module;

3. an exchange unit; 301. a second membrane module; 302. a first exchange chamber; 303. a second exchange chamber;

4. a collection unit; 5. a power unit; 6. a temperature control unit; 61. heating sheet; 62. a cooling sheet; 63. a temperature detecting member; 7. a mixing module;

8. a microscopic observation module; 801. an objective table; 802. a frame; 803. an objective lens; 804. an eyepiece; 805. a first mechanical rail; 806. a second mechanical rail; 807. a camera; 808. a connection part; 809. an autosampler; 810. a sample injection track;

9. a sterile control module; 901. a sterile working chamber; 902. an air inlet fan; 903. a filter assembly; 904. a sterilization assembly;

11. and a fluid supplementing unit.

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments of the present invention. The components of the embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.

Thus, the following detailed description of the embodiments of the invention, as presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

It should be noted that: like reference numerals and letters denote like items in the following figures, and thus once an item is defined in one figure, no further definition or explanation thereof is necessary in the following figures.

In the description of the present invention, it should be noted that, directions or positional relationships indicated by terms such as "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., are directions or positional relationships based on those shown in the drawings, or are directions or positional relationships conventionally put in use of the inventive product, are merely for convenience of describing the present invention and simplifying the description, and are not indicative or implying that the apparatus or element to be referred to must have a specific direction, be configured and operated in a specific direction, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," "third," and the like are used merely to distinguish between descriptions and should not be construed as indicating or implying relative importance. In the description of the present invention, unless otherwise indicated, the meaning of "a plurality" is two or more.

In the description of the present invention, it should also be noted that, unless explicitly specified and limited otherwise, the terms "disposed", "connected" and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected or integrally connected; either mechanically or electrically. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.

In the present invention, unless expressly stated or limited otherwise, a first feature "above" or "below" a second feature may include both the first and second features being in direct contact, as well as the first and second features not being in direct contact but being in contact with each other through additional features therebetween. Moreover, a first feature being "above," "over" and "on" a second feature includes the first feature being directly above and obliquely above the second feature, or simply indicating that the first feature is higher in level than the second feature. The first feature being "under", "below" and "beneath" the second feature includes the first feature being directly under and obliquely below the second feature, or simply means that the first feature is less level than the second feature.

Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative only and are not to be construed as limiting the invention.

Example 1

As shown in fig. 1, the present embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, a collection unit 4 and a power unit 5.

At least one of the premixing unit 1, the culture chamber 2 of the culture unit and the collecting unit 4 is disposable consumable, and is treated as waste after being used once, so that sanitary conditions in the perfusion culture process can be ensured, and normal perfusion culture is prevented from being influenced due to pollution of the premixing unit 1, the culture chamber 2 and the collecting unit 4.

The premix unit 1 comprises a premix chamber 101, a fluid replacement chamber 104 and a gas premix control unit 105, wherein the nutrient solution contains nutrient substances required for the growth of cultures such as culture medium, factors, drugs and enzymes. The fluid replacement chamber 104 is communicated with the premixing chamber 101, the fluid replacement chamber 104 is stored in the nutrient solution, and the fluid replacement chamber 104 is used for conveying the nutrient solution into the premixing chamber 101. A power unit 5 can be arranged between the fluid infusion chamber 104 and the premixing chamber 101 according to the requirement, and the power unit 5 is used for conveying the nutrient solution in the fluid infusion chamber 104 into the premixing chamber 101 according to the requirement of use.

The gas premix control unit 105 communicates with the premix chamber 101 for delivering gases, including in particular oxygen, carbon dioxide, nitrogen, etc., into the premix chamber 101. Nutrient solution and gas are added to the premixing chamber 101 through the fluid supplementing chamber 104 and the gas premixing control unit 105, and the introduced nutrient solution and gas are mixed in the premixing chamber 101 to form a culture solution. The introduced oxygen can increase the dissolved oxygen amount of the culture solution, so that the dissolved oxygen amount can meet the growth requirement of the culture; the introduced carbon dioxide reduces and adjusts the pH value of the culture solution so as to meet the growth requirement of the culture. Preferably, in this embodiment, the number of the premixing chambers 101 and the number of the gas premixing control units 105 are one, and in other embodiments, the number of the premixing chambers 101 and the number of the gas premixing control units 105 are one, and the gas premixing control units 105 are simultaneously communicated with the plurality of premixing chambers 101, so that the gas concentration in the plurality of premixing chambers 101, in particular the dissolved oxygen concentration of the culture solution in the premixing chambers 101, can be controlled.

Preferably, the premixing unit 1 further comprises a pH detecting member 102, an oxygen dissolving detecting member 103 and a signal detector, wherein the pH detecting member 102 and the oxygen dissolving detecting member 103 are arranged in the premixing chamber 101, the signal detector can sense signals of the pH detecting member 102 and the oxygen dissolving detecting member 103, and the sensed signals are calculated and analyzed to obtain the pH value and the oxygen dissolving amount of the culture solution in the premixing chamber 101. With the change of the pH value and the dissolved oxygen amount in the premixing chamber 101, the signals detected by the pH detecting member 102 and the dissolved oxygen detecting member 103 will change, specifically, the optical signals of the pH detecting member 102 and the dissolved oxygen detecting member 103 will change, and the signal detector will periodically receive the optical signals of the surfaces of the pH detecting member 102 and the dissolved oxygen detecting member 103 to obtain data feedback, and the pH value and the dissolved oxygen amount are calculated according to the fed back data. The signal detector can feed back the detected information to the fluid infusion chamber 104 and the gas premixing control unit 105, and if the dissolved oxygen is too high, the fluid infusion chamber 104 is enabled to convey more nutrient solution into the premixing chamber 101 for dilution so as to reduce the dissolved oxygen of the culture solution in the premixing chamber 101; if the dissolved oxygen is too low, the gas premix control unit 105 is caused to introduce oxygen into the premix chamber 101 to increase the dissolved oxygen of the culture solution in the premix chamber 101. If the pH value is too high, the gas premixing control unit 105 is controlled to introduce carbon dioxide so as to reduce the pH value of the culture solution in the premixing chamber 101; if the pH value is too low, the fluid replacement chamber 104 is controlled to convey more nutrient fluid into the premixing chamber 101 for dilution or to introduce lye into the premixing chamber 101 so as to increase the pH value of the culture fluid in the premixing chamber 101.

Specifically, the pH detecting member 102 is a pH electrode sheet, the dissolved oxygen detecting member 103 is a dissolved oxygen electrode sheet, and both are patch materials; the pH electrode plate is provided on the inner wall of the premix chamber 101 or within the premix chamber 101, and the dissolved oxygen detector 103 is provided on the inner wall of the premix chamber 101 or within the premix chamber 101.

The premixing chamber 101 is further provided with an exhaust port, which can exhaust the gas in the premixing chamber 101, so as to ensure the stability of the air pressure of the premixing chamber 101. The premixing chamber 101 is provided with a stirring assembly which can stir the liquid in the premixing chamber 101 to be uniformly mixed and fully immerse the gas in the liquid. The premixing chamber 101 is also provided with an alkali adding port, and alkali liquor can be added into the premixing chamber 101 through the alkali adding port so as to adjust the pH value of the culture solution in the premixing chamber 101.

The nutrient solution and oxygen are premixed in the premixing chamber 101 to form a culture solution, and the culture solution formed by mixing the nutrient solution and the oxygen is more suitable for the growth of the culture by controlling corresponding variables (such as pH value and dissolved oxygen amount), so that nutrients required for the growth of the culture can be sufficiently provided for the culture. Compared with the prior art, the method has the advantages that nutrient solution, oxygen and the like are directly introduced into the culture chamber, so that complex pipelines, valves and detection parts are saved, and the system cost is reduced.

The culture unit comprises a culture chamber 2, a first membrane assembly 203 is arranged in the culture chamber 2, and the first membrane assembly 203 divides the culture chamber 2 into a first culture cavity 205 and a second culture cavity 206. The first culture cavity 205 is communicated with the premixing cavity 101, the premixing cavity 101 can convey culture solution to the first culture cavity 205, and the culture solution in the first culture cavity 205 can flow back into the premixing cavity 101, specifically, the first culture cavity 205 is provided with a liquid inlet and a liquid outlet, and the liquid inlet and the liquid outlet are communicated with the premixing cavity 101. Preferably, a power unit 5 is disposed between the liquid inlet and the premix chamber 101, and the power unit 5 is used to convey the culture liquid in the premix chamber 101 into the first culture chamber 205. Preferably, in this embodiment, the culture chambers 2 are one, i.e. one premix chamber 101 corresponds to one culture chamber. In other embodiments, the culture chambers 2 are multiple, and the culture chambers 2 are all communicated with the premixing chamber 101, i.e. the culture chambers 2 are connected in parallel, which can culture multiple cultures simultaneously, so that consistency of the cultures can be ensured in the subsequent test process, and accuracy of comparison of test results can be ensured.

The collecting unit 4 is communicated with the liquid outlet, the collecting unit 4 is used for collecting the culture solution discharged by the first culture cavity 205, and specifically, the in-vitro life support perfusion culture system further comprises a three-way valve 204, and three interfaces of the three-way valve 204 are respectively communicated with the liquid outlet, the collecting unit 4 and the premixing chamber 101 through pipelines. The switching of the three-way valve can determine whether the culture solution in the first culture chamber 205 is directly discharged into the collecting unit 4 or discharged into the premixing chamber 101 for recycling.

The culture liquid in the first culture chamber 205 can be transferred to the premixing chamber 101 or the collecting unit 4 via the three-way valve 204. Preferably, a power unit 5 is provided between the three-way valve 204 and the liquid discharge port, and the culture liquid in the first culture chamber 205 is fed into the premixing chamber 101 or the collecting unit 4 through the three-way valve 204 by using the power unit 5

The second culture chamber 206 is used to hold a culture and to culture the culture. After the culture solution in the premixing chamber 101 is conveyed into the first culture cavity 205, components required by the growth of the culture in the culture solution in the first culture cavity 205 permeate into the second culture cavity 206 through the first membrane module 203 for the absorption and utilization of the culture; metabolic waste from the growth of the culture may permeate through the first membrane module 203 into the first culture chamber 205. When the concentration of metabolic waste in the culture solution in the first culture cavity 205 is low, the culture solution in the first culture cavity 205 flows back into the premixing cavity 101 through the three-way valve 204, is mixed with gas in the premixing cavity 101, and is then conveyed into the first culture cavity 205; when the metabolic waste concentration of the culture solution in the first culture cavity 205 is higher or the concentration of the components needed by the culture is lower, the three-way valve 204 is controlled, so that the culture solution flowing out of the first culture cavity 205 is discharged into the collecting unit 4 through the three-way valve 204, and then the nutrient solution is supplemented into the premixing cavity 101 by the nutrient solution supplementing cavity 104.

In addition, the first membrane module 203 can prevent the culture from being directly washed by the circulating culture solution, and can also entrap the cells of the culture, so that the single dispersed cells of the culture are not washed away, and the system can be applied to the culture of more cultures.

The culture chamber 2 is provided with a sample inlet 201 and a sample inlet 202, and the sample inlet 201 and the sample inlet 202 are specifically arranged on the second culture chamber 206. The culture may be added to the second culture chamber 206 through the sample addition port 201. And growth factors, medicines and the like can be added into the second culture cavity 206 to promote or inhibit the growth of the culture in the culture process according to experimental requirements, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into the system after the culture is completed to collect, treat or characterize the culture. A sample of the culture or broth may be taken for analysis during the culturing process via sampling port 202.

Because the consumption speed of each component in the culture solution is different in the culture process, one or more components can be directionally added through the fluid infusion chamber 104, so that the balance of each component is ensured, the whole fluid exchange step is reduced, the cost is reduced, and the pollution probability of the culture is reduced. The fluid replacement chamber 104 supplements the premix chamber 101 with fluid, and then the premix chamber 101 conveys the nutrient solution to the culture chamber 2; thus, the concentration and composition of the nutrient solution in the fluid-filled chamber 104 need not be formulated to be the same as the concentration and composition of the culture solution required for the culture, and the volume of the fluid-filled chamber 104 can be made smaller. Meanwhile, the perfusion rate of the pre-mixing chamber 101 can be higher without worrying about excessively fast consumption of the culture solution due to the increase of the perfusion rate, improving the culture efficiency and increasing the kinds of cultures that can be cultured.

Specifically, the concentration of the culture medium component flowing out of the culture chamber 2 can be detected, and the culture medium component can be purposefully replenished through the replenishing chamber 104. The concentration detection can be sampling detection or in-situ detection (such as infrared spectrum technology and fluorescence detection technology). Preferably, this embodiment employs fluorescence detection techniques. The system may also be provided with an automatic sample addition module that automatically adds nutrients to the fluid replacement chamber 104 based on the concentration measurements described above.

Preferably, the culture unit further comprises a temperature control unit 6. The temperature control unit 6 is used for controlling the temperature of the culture chamber 2, and specifically can heat the premixing chamber 101 and the culture chamber 2 to enable the temperature of the culture solution in the premixing chamber 101 and the temperature of the culture solution in the culture chamber 2 to rise to the physiological temperature suitable for the growth of the culture so as to culture the culture; after the culture is finished, when the matrigel is needed to be removed and the culture is collected, the temperature control unit 6 can control the temperature of the culture solution in the culture chamber 2 to a first preset temperature, wherein the first preset temperature is the liquefaction temperature of the culture supporting structure, and the culture can be taken out through the sampling port 202 after the culture supporting structure is liquefied. The culture support structure forms a network support for the culture, facilitating 3D growth of the culture. Culture support structures include, but are not limited to, matrigel (2-8 degrees liquefaction), synthetic gel (25 degrees liquefaction at room temperature). Specifically, the temperature control unit 6 includes heating component, refrigeration component and temperature control module, and heating component and refrigeration component are all connected with temperature control module electricity, and temperature control module is used for controlling heating component heating culture cavity 2, still is used for controlling refrigeration component cooling culture cavity 2.

Heating components and refrigerating components are arranged on the outer walls of the premixing chamber 101 and the culturing chamber 2, the heating components comprise a plurality of heating sheets 61, and the heating sheets 61 are arranged on the outer wall of the culturing chamber 2. The heating sheet 61 may be a transparent heating plate, a metal heating plate, a semiconductor heating plate, a blanket, a sheet, or the like. The refrigerating assembly comprises a plurality of refrigerating sheets 62, the refrigerating sheets 62 are arranged on the outer wall of the culture chamber 2, and the refrigerating sheets 62 can be semiconductor refrigerating sheets 62 or other refrigerating materials.

The heating plate 61 and the cooling plate 62 may be located at different positions of the culture chamber 2, for example: as shown in fig. 2, the heating plate 61 is positioned at the bottom of the culture chamber 2, and the cooling plate 62 is positioned at the side of the culture chamber 2; alternatively, as shown in FIG. 3, the heating sheet 61 is located on the side of the culture chamber 2, and the cooling sheet 62 is located on the bottom of the culture chamber 2. Of course, the heating plate 61 and the cooling plate 62 may be located at the same position of the culture chamber 2, for example, both may be located at the same side or bottom of the culture chamber 2.

Preferably, in this embodiment, as shown in FIG. 4, the heating sheet 61 and the cooling sheet 62 are located on the same side or bottom wall of the culture chamber 2. Specifically, the plurality of heating sheets 61 are arranged at intervals, the plurality of cooling sheets 62 are arranged at intervals, one cooling sheet 62 is arranged between two adjacent heating sheets 61, one heating sheet 61 is arranged between two adjacent cooling sheets 62, moreover, the heating sheets 61 are in close contact with the adjacent cooling sheets 62, and the cooling sheets 62 are in close contact with the adjacent heating sheets 61, so that energy of the heating sheets 61 can be transferred to the cooling sheets 62. The heating sheet 61 and the cooling sheet 62 are both made of a material with good heat conductivity, and are structurally processed into an inlaid structure, so that the heating sheet 61 is inlaid between the cooling sheets 62, and the cooling sheet 62 is inlaid between the heating sheets 61.

When the temperature of the culture chamber 2 needs to be raised, the heating plate 61 is heated to a specified temperature under the control of the temperature control module, and the cooling plate 62 is heated while the culture chamber 2 is heated, and then the temperature of the cooling plate 62 is quickly raised to the specified temperature, and the culture chamber 2 is heated by the heating plate 61 and the cooling plate 62 together, because the cooling plate 62 is made of a heat-conducting material, and the heat of the heating plate 61 can be transferred to the cooling plate 62. When the culture chamber 2 needs to be cooled down, the cooling plate 62 is cooled down to a specified temperature under the control of the temperature control module, as the heating plate 61 is made of a heat conducting material, and the heat of the heating plate 61 can be transferred to the cooling plate 62, the cooling plate 62 cools down the culture chamber 2 and simultaneously cools down the heating plate 61 to enable the temperature to be rapidly cooled down to the specified temperature, and then the cooling plate 62 and the heating plate 61 cool down the culture chamber 2 together.

The heating plates 61 and the cooling plates 62 are arranged in a staggered manner, and can transfer heat, so that the heating plates and the cooling plates can jointly heat and cool the culture chamber 2, the heat exchange area is increased, and the heat exchange efficiency is improved.

The temperature control unit 6 further comprises a temperature detecting member 63, which may specifically be a temperature sensor. The temperature detection piece 63 is electrically connected with the temperature control module, and the temperature detection piece 63 can be arranged on the inner wall of the culture chamber 2 or on the outer wall of the culture chamber 2, and is used for detecting the temperature of the culture solution in the culture chamber and feeding back the detection value to the temperature control module, and the temperature control module compares the temperature value detected by the temperature detection piece 63 with a set value actually required to control the heating component to heat the temperature culture chamber 2 or control the refrigerating component to cool the culture chamber 2.

The heating unit, cooling unit and temperature detecting unit 63 of the premix chamber 101 are constructed and arranged in the same manner as those of the incubation chamber 2, and will not be described in detail herein.

Optionally, with continued reference to fig. 1, the culture unit further comprises a mixing module 7, the mixing module 7 being in particular a mechanical shaking mechanism for shaking the culture liquid in the culture chamber 2, in particular by shaking the culture chamber 2, to shake the culture liquid in the culture chamber 2, providing a more sufficient exchange opportunity for the culture liquid and the culture. The shaking amplitude and speed of the mixing module 7 may be matched to the speed of feeding the culture medium into the culture chamber 2, for example, a higher feeding speed of the culture medium may promote exchange of the culture medium with the culture medium, but may cause excessive consumption of the culture medium. The lower culture solution conveying speed is matched with large-amplitude mechanical shaking, so that the purposes of updating the culture solution, increasing the exchange opportunity of the nutrient solution and the culture and promoting the exchange of nutrient substances can be simultaneously achieved.

Optionally, as shown in fig. 5 and 6, the culture unit further comprises a microscopic observation module 8 comprising a stage 801 and an observation assembly, the culture chamber 2 being placed on the stage 801, in particular the culture chamber 2 being placed on the mixing module 7, the mixing module 7 being placed on the stage 801. The stage 801 is capable of placing at least one culture chamber 2, in this embodiment, a placing station is provided on the stage 801, and the mixing module 7 is placed on the placing station; in other embodiments, a plurality of placement stations are provided on the stage 801, each of which can place one blending module 7.

The viewing assembly is for viewing the culture in the culture chamber 2, in particular the viewing assembly comprises an objective 803, an eyepiece 804 and a camera 807, in particular a CCD camera. The microscopic observation module 8 further includes a frame 802, and an objective 803 is provided on the frame 802 so as to be movable in a vertical direction, and an eyepiece 804 is also provided on the frame 802, through which eyepiece 804 the culture in the culture chamber 2 can be observed.

The frame 802 is provided with a first mechanical rail 805, and a second mechanical rail 806 is slidably provided on the first mechanical rail 805, where the first mechanical rail 805 and the second mechanical rail 806 are disposed perpendicular to each other and are both located in a horizontal plane. The stage 801 is slidably disposed on the second mechanical rail 806, and adjustment of any position of the stage 801 in a horizontal plane can be achieved by sliding the stage 801 relative to the second mechanical rail 806 and sliding the second mechanical rail 806 relative to the first mechanical rail 805 so that the objective 803 is facing the observation chamber, and the culture is observed by focusing the eyepiece 804 on the culture in the culture chamber 2. If image data is to be recorded during the observation, the camera 807 may be used to take a photograph of the recorded image data.

Preferably, as shown in fig. 7, the microscopic observation module 8 further includes an auto-sampler 809 and a sampling rail 810, wherein the auto-sampler 810 is disposed on the frame 802, the auto-sampler 809 is slidingly disposed on the sampling rail 810, and the auto-sampler 809 is driven by the sampling driving member to slide on the sampling rail 810 to perform sampling work. The automatic sampler 809 is mainly used for adding samples when the culture, reagent or medicine and the like are needed to be added in the culture chamber 2, so that the manual sample adding is avoided, the sanitary condition of the culture chamber 2 is ensured, and the culture is not easy to be polluted.

Optionally, as shown in fig. 8, the in vitro life support perfusion culture system further comprises a sterile control module 9, which comprises a sterile working chamber 901, a filtering component 903 and a sterilizing component 904, at least the culture chamber 2 is arranged in the sterile working chamber 901, and in this embodiment, the premixing unit 1, the culture unit, the collecting unit 4, the power unit 5 and the like are all arranged in the sterile working chamber 901, i.e. the part of the culture system of the culture except the sterile control module 9 is located in the sterile working chamber 901.

The aseptic working chamber 901 is provided with the vent, and filter component 903 sets up in the vent department, and is provided with air inlet fan 902 on the vent, lets in gas in to aseptic working chamber 901 through air inlet fan 902, and filter component 903 is used for filtering the gas that lets in aseptic working chamber 901, makes dust magazine etc. unable entering aseptic working chamber 901. The sterilization component 904 is used for sterilizing the aseptic working chamber 901, specifically, the sterilization component 904 is an ultraviolet lamp, and is arranged at the air inlet, the sterilization component 904 can sterilize the aseptic working chamber 901 and sterilize the gas introduced into the aseptic working chamber 901, and the sterilization component 904 can prevent the gas with bacteria from entering the aseptic working chamber 901.

The embodiment also discloses a control method of the in-vitro life support perfusion culture system, which aims at the in-vitro life support perfusion culture system and comprises the following steps:

the concentration of each component required by the culture in the culture solution in the culture chamber 2 is detected and/or the growth condition of the culture is determined, the speed of delivering the culture solution to the culture chamber 2 by the premixing chamber 101 is controlled according to the detected concentration and/or the determined growth condition of the culture, and the speed of delivering the nutrient solution to the premixing chamber 1 by the fluid supplementing chamber 104 and the speed of delivering the gas to the premixing chamber 101 by the gas premixing control unit 105 are also controlled.

Example two

The embodiment discloses an in-vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, a collecting unit 4 and a power unit 5.

The in vitro life support perfusion culture system in this embodiment is basically the same as the in vitro life support perfusion culture system in the first embodiment, except that the culture unit does not include the temperature control unit 6 and the mixing unit, and the culture unit in this embodiment further includes a mixing temperature control module for controlling the temperature of the culture chamber 2 and shaking the culture solution in the culture chamber 2.

The mixing temperature control module comprises a mechanical shaking mechanism and a temperature control assembly, and the mechanical shaking mechanism is provided with a mounting surface. The temperature control component is used for controlling the temperature of the culture chamber 2, and specifically can heat the premixing chamber 101 and the culture chamber 2 to ensure that the temperature of the culture solution in the premixing chamber 101 and the temperature of the culture solution in the culture chamber 2 are raised to the physiological temperature suitable for the growth of the culture so as to culture the culture; after the culture is completed, when the matrigel removal is required and the culture is collected, the temperature control assembly can lower the culture solution in the culture chamber 2 to the matrigel melting temperature in the culture solution so as to collect the culture. Specifically, the temperature control assembly comprises a heating assembly, a refrigerating assembly and a temperature control module, wherein the heating assembly and the refrigerating assembly are electrically connected with the temperature control module, and the temperature control module is used for controlling the heating assembly to heat the culture chamber 2 and controlling the refrigerating assembly to cool the culture chamber 2.

The heating assembly comprises a plurality of heating plates 61, the heating plates 61 being arranged on the outer wall of the culture chamber 2. The heating sheet 61 may be a transparent heating plate, a metal heating plate, a semiconductor heating plate, a blanket, a sheet, or the like. The refrigerating assembly comprises a plurality of refrigerating sheets 62, the refrigerating sheets 62 are arranged on the outer wall of the culture chamber 2, and the refrigerating sheets 62 can be semiconductor refrigerating sheets 62 or other refrigerating materials. Specifically, the plurality of heating sheets 61 are arranged at intervals, the plurality of cooling sheets 62 are arranged at intervals, one cooling sheet 62 is arranged between two adjacent heating sheets 61, one heating sheet 61 is arranged between two adjacent cooling sheets 62, moreover, the heating sheets 61 are in close contact with the adjacent cooling sheets 62, and the cooling sheets 62 are in close contact with the adjacent heating sheets 61, so that energy of the heating sheets 61 can be transferred to the cooling sheets 62. The heating sheet 61 and the cooling sheet 62 are both made of a material with good heat conductivity, and are structurally processed into an inlaid structure, so that the heating sheet 61 is inlaid between the cooling sheets 62, and the cooling sheet 62 is inlaid between the heating sheets 61. The heating sheet 61 and the cooling sheet 62 are sequentially arranged on the mounting surface and are mainly used for heating or cooling the bottom surface of the culture chamber 2, so as to control the temperature of the culture solution in the culture chamber 2.

The temperature control assembly further comprises a temperature sensing member 63, which may specifically be a temperature sensor. The temperature detection piece 63 is electrically connected with the temperature control module, and the temperature detection piece 63 can be arranged on the inner wall of the culture chamber 2 or on the outer wall of the culture chamber 2 for detecting the temperature of the culture solution in the culture chamber and feeding back the detection value to the temperature control module, and the temperature control module compares the temperature value detected by the temperature detection piece 63 with a set value actually required to control the heating component to heat the temperature culture chamber 2 or control the refrigerating component to cool the culture chamber 2.

Example III

The in vitro life support perfusion culture system in this embodiment is basically the same as that in embodiment one, except that the culture unit in this embodiment does not include the mixing module 7, and the structure of the microscopic observation unit is also different. As shown in fig. 9, the microscopic observation module 8 in the present embodiment includes a stage 801, an observation assembly, and a frame 802, the culture chamber 2 is placed on the stage 801, the stage 801 is swingably provided on the frame 802, and the swing stage 801 can shake the culture liquid in the culture chamber 2.

The culture chamber 2 is placed on the stage 801, and the culture chamber 2 is placed on the stage 801. The stage 801 is capable of placing at least one culture chamber 2, and in this embodiment, the stage 801 is provided with a placing station, and the culture chamber 2 is placed on the placing station; in other embodiments, a plurality of placement stations are provided on stage 801, each of which can place one culture chamber 2.

The viewing assembly is for viewing the culture in the culture chamber 2, in particular the viewing assembly comprises an objective 803, an eyepiece 804 and a camera 807, in particular a CCD camera 807. The microscopic observation module 8 further includes a frame 802, and an objective 803 is provided on the frame 802 so as to be movable in a vertical direction, and an eyepiece 804 is also provided on the frame 802, through which eyepiece 804 the culture in the culture chamber 2 can be observed.

The frame 802 is provided with a first mechanical rail 805, and a second mechanical rail 806 is slidably provided on the first mechanical rail 805, where the first mechanical rail 805 and the second mechanical rail 806 are disposed perpendicular to each other and are both located in a horizontal plane. The connection portion 808 is mounted on the bottom of the stage 801, a rotation shaft is provided on the connection portion 808, the stage 801 is rotatably connected to the rotation shaft, the stage 801 can swing with respect to the connection portion 808, and the connection portion 808 is slidably provided on the second mechanical rail 806 and can slide along the longitudinal direction of the second mechanical rail 806. The microscopic observation module 8 further comprises a driving member, the output end of which is connected to the stage 801 for driving the stage 801 to shake so as to shake the culture solution in the culture chamber 2. Providing more sufficient exchange opportunities for culture fluids and cultures. The shaking amplitude and speed of stage 801 may be matched to the rate of transfer of culture medium into culture chamber 2, e.g., a higher culture medium transfer rate may facilitate exchange of culture medium with the culture, but may result in excessive consumption of culture medium. The lower culture solution conveying speed is matched with large-amplitude mechanical shaking, so that the purposes of updating the culture solution, increasing the exchange opportunity of the nutrient solution and the culture and promoting the exchange of nutrient substances can be simultaneously achieved.

Adjustment of any position of the stage 801 in the horizontal plane can be achieved by sliding the stage 801 relative to the second mechanical rail 806 and the second mechanical rail 806 relative to the first mechanical rail 805 so that the objective 803 is facing the viewing chamber and the culture is viewed using the eyepiece 804 to focus on the culture in the culture chamber 2. If image data is to be recorded during the observation, the camera 807 may be used to take a photograph of the recorded image data. The stage 801 may be shaken during the observation to bring the culture into a better view for ease of observation.

Preferably, as shown in fig. 10, the microscopic observation module 8 further includes an auto-sampler 809 and a sampling rail 810, wherein the auto-sampler 810 is disposed on the frame 802, the auto-sampler 809 is slidingly disposed on the sampling rail 810, and the auto-sampler 809 is driven by the sampling driving member to slide on the sampling rail 810 to perform sampling work. The automatic sampler 809 is mainly used for adding samples when the culture, reagent or medicine and the like are needed to be added in the culture chamber 2, so that the manual sample adding is avoided, the sanitary condition of the culture chamber 2 is ensured, and the culture is not easy to be polluted.

Example IV

As shown in fig. 11, the present embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, a collection unit 4 and a power unit 5.

The structure of the pre-mix unit 1 is the same as that of the pre-mix unit 1 of the first embodiment and will not be described in detail here.

The culture unit comprises a culture chamber 2, a first membrane assembly 203 is arranged in the culture chamber 2, and the first membrane assembly 203 divides the culture chamber 2 into a first culture cavity 205 and a second culture cavity 206. The first culture chamber 205 is in communication with the premix chamber 101, the premix chamber 101 is capable of delivering culture fluid to the first culture chamber 205, the second culture chamber 206 is in communication with the premix chamber 101, and culture fluid in the second culture chamber 206 is capable of flowing back into the premix chamber 101. Specifically, the first culture chamber 205 is provided with a liquid inlet, the second culture chamber 206 is provided with a liquid outlet, and both the liquid inlet and the liquid outlet are communicated with the premixing chamber 101. The culture solution in the premixing chamber 101 can be conveyed into the first culture chamber 205, and whether the power unit 5 is arranged between the premixing chamber 101 and the liquid inlet or not can be determined according to the relative arrangement position of the liquid inlets of the premixing chamber 101 and the first culture chamber 205 and the conveying speed and the conveying amount of the culture solution, and the culture solution in the premixing chamber 101 is conveyed into the first culture chamber 205 by using the power unit 5.

The collecting unit 4 is communicated with the liquid outlet, the collecting unit 4 is used for collecting the culture solution discharged by the second culture cavity 206, and specifically, the in-vitro life support perfusion culture system further comprises a three-way valve 204, and three interfaces of the three-way valve 204 are respectively communicated with the liquid outlet, the collecting unit 4 and the premixing chamber 101 through pipelines. The switching of the three-way valve 204 can determine whether the culture solution in the first culture chamber 205 is directly discharged into the collecting unit 4 or discharged into the premixing chamber 101 for recycling.

The culture liquid in the second culture chamber 206 can be transferred to the premixing chamber 101 or the collecting unit 4 via the three-way valve 204. According to the relative positions of the three-way valve 204 and the liquid outlet of the second culture chamber 206 and the requirements of the conveying speed and the conveying amount of the culture liquid, it can be determined whether a power unit 5 is arranged between the three-way valve 204 and the liquid outlet, and the culture liquid in the second culture chamber 206 is conveyed into the premixing chamber 101 or the collecting unit 4 by the power unit 5 through the three-way valve 204

Preferably, in this embodiment, the second culture chamber 206 is used to hold a culture and to culture the culture; after the culture solution in the premixing chamber 101 is conveyed into the first culture cavity 205, components required by growth of the culture in the culture solution in the first culture cavity 205 permeate into the second culture cavity 206 through the first membrane module 203 for absorption and utilization of the culture, and the components can be buffered through the first membrane module 203 to avoid flushing the culture. Furthermore, the perfusion pressure of the nutrient solution can enable the nutrient solution and the culture to be fully exchanged on the surface of the first membrane module 203, so as to promote the absorption of nutrient substances to the greatest extent. Specifically, the culture chamber 2 is provided with a loading port 201 and a sampling port 202, and the loading port 201 and the sampling port 202 are specifically provided on the second culture chamber 206. The culture may be added to the second culture chamber 206 through the sample addition port 201. And growth factors, medicines and the like can be added into the second culture cavity 206 to promote or inhibit the growth of the culture in the culture process according to experimental requirements, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into the system after the culture is completed to collect, treat or characterize the culture. A sample of the culture or broth may be taken for analysis during the culturing process via sampling port 202.

In other embodiments, first culture chamber 205 is used to hold and culture a culture. The perfusion pressure of the culture fluid flow can enable the culture fluid and the culture to be fully exchanged on the surface of the first membrane module 203, so that the absorption of nutrient substances is promoted to the greatest extent. The culture chamber 2 is provided with a loading port 201 and a sampling port 202, and specifically, the loading port 201 and the sampling port 202 are provided specifically on the first culture chamber 205. The culture may be added to the first culture chamber 205 through the loading port 201. And growth factors, medicines and the like can be added into the first culture cavity 205 through the sample adding port 201 to promote or inhibit the growth of the culture according to experimental requirements, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into a system after the culture is completed to collect, treat or characterize the culture. A sample of the culture or broth may be taken for analysis during the culturing process via sampling port 202.

Preferably, the culture unit further includes a temperature control unit 6 and a micro-observation module 8, and the temperature control unit 6 and the micro-observation module 8 are identical in structure to the temperature control unit 6 and the micro-observation module 8 in the first embodiment, and will not be described in detail herein.

Optionally, the in vitro life support perfusion culture system further comprises a sterility control module 9, and the sterility control module 9 has the same structure as the sterility control module 9 in the first embodiment, and will not be described in detail here.

Example five

As shown in fig. 12, the present embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, an exchange unit 3, a fluid supplementing unit 11, a collecting unit 4 and a power unit 5.

At least one of the premixing unit 1, the culturing chamber 2 of the culturing unit, the exchanging unit 3 and the collecting unit 4 is disposable, and is treated as waste after being used once, so that sanitary conditions in the perfusion culturing process can be ensured, and normal perfusion culturing is prevented from being influenced due to pollution of the premixing unit 1, the culturing chamber 2, the exchanging unit 3 and the collecting unit 4.

The premix unit 1 comprises a premix chamber 101 and a gas premix control unit 105, wherein the culture medium contains nutrients required for the growth of the culture such as culture medium, factors, drugs and enzymes. The gas premix control unit 105 communicates with the premix chamber 101 for delivering gases, including in particular oxygen, carbon dioxide, nitrogen, etc., into the premix chamber 101. The premix chamber 101 may receive a nutrient solution in a manner that will not be described in greater detail herein. Gas is added to the premix chamber 101 by the gas premix control unit 105, and the introduced nutrient solution and gas are mixed in the premix chamber 101 to form a culture solution. The introduced oxygen can increase the dissolved oxygen amount of the culture solution, so that the dissolved oxygen amount can meet the growth requirement of the culture; the introduced carbon dioxide reduces and adjusts the pH value of the culture solution so as to meet the growth requirement of the culture. Preferably, in this embodiment, the number of the premixing chambers 101 and the number of the gas premixing control units 105 are one, and in other embodiments, the number of the premixing chambers 101 and the number of the gas premixing control units 105 are one, and the gas premixing control units 105 are simultaneously communicated with the plurality of premixing chambers 101, so that the gas concentration in the plurality of premixing chambers 101, in particular the dissolved oxygen concentration of the culture solution in the premixing chambers 101, can be controlled.

Preferably, the signal detector is capable of sensing the signals of the pH detecting element 102 and the dissolved oxygen detecting element 103, and calculating and analyzing the sensed signals to obtain the pH value and the dissolved oxygen amount of the culture solution in the premixing chamber 101. With the change of the pH value and the dissolved oxygen amount in the premixing chamber 101, the signals detected by the pH detecting member 102 and the dissolved oxygen detecting member 103 will change, specifically, the optical signals of the pH detecting member 102 and the dissolved oxygen detecting member 103 will change, and the signal detector will periodically receive the optical signals of the surfaces of the pH detecting member 102 and the dissolved oxygen detecting member 103 to obtain data feedback, and the pH value and the dissolved oxygen amount are calculated according to the fed back data. The signal detector can feed back the detected information to the gas premix control unit 105, and if the dissolved oxygen is too low, the gas premix control unit 105 is caused to introduce oxygen into the premix chamber 101 to increase the dissolved oxygen of the culture fluid in the premix chamber 101. If the pH is too high, the gas premix control unit 105 is controlled to pass carbon dioxide to reduce the pH of the broth in the premix chamber 101. If the pH value is too low, lye is introduced into the premixing chamber 101 to increase the pH value of the culture solution in the premixing chamber 101.

The culture unit comprises a culture chamber 2, a first membrane assembly 203 is arranged in the culture chamber 2, and the first membrane assembly 203 divides the culture chamber 2 into a first culture chamber 205 and a second culture chamber 206; the first culture cavity 205 is communicated with the premixing cavity 101, the premixing cavity 101 can convey culture solution to the first culture cavity 205, the culture solution in the first culture cavity 205 can flow back into the premixing cavity 101, and specifically, the first culture cavity 205 is provided with a liquid inlet and a liquid outlet, and the liquid inlet is communicated with the premixing cavity 101. The culture solution in the premixing chamber 101 can be conveyed into the first culture chamber 205, and whether the power unit 5 is arranged between the premixing chamber 101 and the liquid inlet or not can be determined according to the relative arrangement position of the liquid inlets of the premixing chamber 101 and the first culture chamber 205 and the conveying speed and the conveying amount of the culture solution, and the culture solution in the premixing chamber 101 is conveyed into the first culture chamber 205 by using the power unit 5.

Preferably, in this embodiment, the culture chambers 2 are one, i.e. one premix chamber 101 corresponds to one culture chamber. In other embodiments, the culture chambers 2 are multiple, and the culture chambers 2 are all communicated with the premixing chamber 101, i.e. the culture chambers 2 are connected in parallel, which can culture multiple cultures simultaneously, so that consistency of the cultures can be ensured in the subsequent test process, and accuracy of comparison of test results can be ensured.

The second culture chamber 206 is used for containing a culture and culturing the culture; after the culture solution in the premixing chamber 101 is conveyed into the first culture cavity 205, components required by the growth of the culture in the culture solution in the first culture cavity 205 permeate into the second culture cavity 206 through the first membrane module 203 for the absorption and utilization of the culture; metabolic waste from the growth of the culture may permeate through the first membrane module 203 into the first culture chamber 205.

The exchange unit 3 comprises an exchange chamber and a second membrane module 301, the exchange chamber comprises a first exchange cavity 302 and a second exchange cavity 303, the second membrane module 301 is arranged between the first exchange cavity 302 and the second exchange cavity 303, and is communicated with the first exchange cavity and the second exchange cavity through the membrane module 301, and the second membrane module 301 is used for intercepting and/or penetrating partial components in the culture solution. Specifically, the second membrane module 301 may trap useful components of the culture solution in the first exchange chamber 302, such as serum, growth factors, enzymes, etc., and metabolites produced by the growth of the culture in the first exchange chamber 302, such as urea, carbon dioxide, etc., may permeate through the membrane module into the second exchange chamber 303 and then be discharged from the second exchange chamber 303.

The exchange unit 3 and the second membrane component 301 are arranged, so that useful components in the culture solution can be intercepted while perfusion culture is realized, and the useful components are reversely flowed into the culture chamber 2 for re-absorption and utilization, thereby improving the utilization rate of the culture solution and reducing the culture cost.

The liquid outlet and the premixing unit 1 are both communicated with the first exchange cavity 302, specifically, a first interface and a second interface are arranged on the first exchange cavity 302, the liquid outlet is communicated with the first interface, and the premixing unit 1 is communicated with the second interface. The culture solution in the first culture cavity 205 can be conveyed into the first exchange cavity 302 through the liquid outlet and the first interface, and whether the power unit 5 is arranged between the first interface and the liquid outlet of the first exchange cavity 302 or not can be determined according to the relative arrangement position of the first interface of the first exchange cavity 302 and the liquid outlet of the first culture cavity 205 and the conveying speed and the conveying amount of the culture solution, and the power unit 5 is used for conveying the culture solution in the first culture cavity 205 into the first exchange cavity 302. The culture fluid in the first exchange chamber 302 can flow into the premixing chamber 101 through the second port, so that useful components in the culture fluid in the first exchange chamber 302 are recycled. Depending on the relative arrangement of the second interface of the first exchange chamber 302 and the premix chamber 101 and the requirements of the transfer rate and transfer volume of the culture solution, it may be decided whether a power unit 5 is arranged between the second interface of the first exchange chamber 302 and the premix chamber 101, and the culture solution in the first exchange chamber 302 is transferred into the premix chamber 101 via the second interface using the power unit 5.

The collection unit 4 communicates with the second exchange chamber 303, and the collection unit 4 is used for collecting the culture solution discharged from the second exchange chamber 303. Specifically, the second exchange chamber 303 is provided with a third interface and a fourth interface, and the fourth interface is communicated with the collecting unit 4. The collection unit 4 communicates with the second exchange chamber 303 for collecting the culture liquid in the second exchange chamber 303. Preferably, in this embodiment, a power unit 5 is disposed between the collection unit 4 and the second exchange chamber 303, and the power unit 5 is used for conveying the culture solution in the second exchange chamber 303 into the collection unit 4. Depending on the relative arrangement of the second exchange chamber 303 and the collecting unit 4 and the requirements of the transport speed and transport amount of the culture liquid, it may be decided whether or not a power unit 5 is provided between the second exchange chamber 303 and the collecting unit 4, and the power unit 5 is used to transport the culture liquid in the second exchange chamber 303 to the collecting unit 4.

The fluid infusion unit 11 communicates with the second exchange chamber 303, in particular the third interface communicates with the fluid infusion unit 11. The fluid replacement unit 11 is configured to deliver a supplemental nutrient solution to the second exchange chamber 303, where at least a portion of components in the supplemental nutrient solution in the second exchange chamber 303 can permeate through the membrane assembly into the first exchange chamber 302. A power unit 5 is arranged between the fluid infusion unit 11 and the second exchange cavity 303, and whether the power unit 5 is arranged between the fluid infusion unit 11 and the second exchange cavity 303 can be determined according to the relative arrangement position of the second exchange cavity 303 and the fluid infusion unit 11 and the requirements of the delivery speed and the delivery quantity of the supplementary nutrient fluid, and the supplementary nutrient fluid is delivered by using the power unit 5. The concentration of the supplementary nutrient solution is greater than that of the culture solution in the first exchange cavity 302, and the concentration difference between the two can drive components in the supplementary nutrient solution to move into the first exchange cavity 302 through the membrane component, and then the components are conveyed into the premixing cavity 101 to supplement the nutrient solution, so that the nutrient solution and the gas are mixed in the premixing cavity 101 to form the culture solution.

Because the consumption speed of each component in the culture solution is different in the culture process, one or more components can be directionally added through the fluid infusion unit 11 according to the scheme of the embodiment, so that the balance of each component is ensured, the whole fluid exchange step is reduced, the cost is reduced, and the pollution probability of the culture is reduced. The fluid supplementing unit 11 supplements the second exchange cavity 303 with fluid, and then the first exchange cavity 302 conveys the nutrient solution to the culture cavity 2; therefore, the concentration and the composition of the nutrient solution in the fluid infusion unit 11 do not have to be formulated to be the same as those of the culture solution required for the culture, and the volume of the fluid infusion unit 11 can be made smaller. Meanwhile, the perfusion rate of the pre-mixing chamber 101 can be higher without worrying about excessively fast consumption of the culture solution due to the increase of the perfusion rate, improving the culture efficiency and increasing the kinds of cultures that can be cultured.

In particular, the concentration of the culture medium components flowing out of the culture chamber 2 can be detected and purposefully replenished by the replenishing unit 11. The concentration detection can be sampling detection or in-situ detection (such as infrared spectrum technology and fluorescence detection technology). Preferably, this embodiment employs fluorescence detection techniques. The system may also be provided with an automatic sample adding module for automatically adding nutritional components to the fluid replacement unit 11 based on the above concentration detection results.

Example six

As shown in fig. 13, the present embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, an exchange unit 3, a fluid supplementing unit 11, a collecting unit 4 and a power unit 5.

At least one of the premixing unit 1, the culture chamber 2 of the culture unit and the collecting unit 4 is disposable, and is treated as waste after being used once, so that sanitary conditions in the perfusion culture process can be ensured, and normal perfusion culture is prevented from being influenced due to pollution of the premixing unit 1, the culture chamber 2 and the collecting unit 4.

The structure of the pre-mix unit 1 is the same as that of the pre-mix unit 1 in embodiment five and will not be described in detail here.

The culture unit comprises a culture chamber 2, a first membrane assembly 203 is arranged in the culture chamber 2, and the first membrane assembly 203 divides the culture chamber 2 into a first culture chamber 205 and a second culture chamber 206;

The first culture chamber 205 is in communication with the premix chamber 101, the premix chamber 101 is capable of delivering culture fluid to the first culture chamber 205, the second culture chamber 206 is in communication with the premix chamber 101, and culture fluid in the second culture chamber 206 is capable of flowing back into the premix chamber 101. Specifically, the first culture chamber 205 is provided with a liquid inlet, the second culture chamber 206 is provided with a liquid outlet, and the liquid inlet premix chamber 101 is communicated. The culture solution in the premixing chamber 101 can be conveyed into the first culture chamber 205, and whether the power unit 5 is arranged between the premixing chamber 101 and the liquid inlet or not can be determined according to the relative arrangement position of the liquid inlets of the premixing chamber 101 and the first culture chamber 205 and the conveying speed and the conveying amount of the culture solution, and the culture solution in the premixing chamber 101 is conveyed into the first culture chamber 205 by using the power unit 5. Preferably, in this embodiment, the culture chambers 2 are one, i.e. one premix chamber 101 corresponds to one culture chamber. In other embodiments, the culture chambers 2 are multiple, and the culture chambers 2 are all communicated with the premixing chamber 101, i.e. the culture chambers 2 are connected in parallel, which can culture multiple cultures simultaneously, so that consistency of the cultures can be ensured in the subsequent test process, and accuracy of comparison of test results can be ensured.

Preferably, in this embodiment, the first culture chamber 205 is used to hold a culture and to culture the culture. The perfusion pressure of the culture fluid flow can enable the culture fluid and the culture to be fully exchanged on the surface of the first membrane module 203, so that the absorption of nutrient substances is promoted to the greatest extent. The culture chamber 2 is provided with a loading port 201 and a sampling port 202, and specifically, the loading port 201 and the sampling port 202 are provided specifically on the first culture chamber 205. The culture may be added to the first culture chamber 205 through the loading port 201. And growth factors, medicines and the like can be added into the first culture cavity 205 through the sample adding port 201 to promote or inhibit the growth of the culture according to experimental requirements, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into a system after the culture is completed to collect, treat or characterize the culture. A sample of the culture or broth may be taken for analysis during the culturing process via sampling port 202.

In other embodiments, second culture chamber 206 is used to hold and culture a culture; after the culture solution in the premixing chamber 101 is conveyed into the first culture cavity 205, components required by growth of the culture in the culture solution in the first culture cavity 205 permeate into the second culture cavity 206 through the first membrane module 203 for absorption and utilization of the culture, and the components can be buffered through the first membrane module 203 to avoid flushing the culture. Furthermore, the perfusion pressure of the nutrient solution can enable the nutrient solution and the culture to be fully exchanged on the surface of the first membrane module 203, so as to promote the absorption of nutrient substances to the greatest extent. The culture chamber 2 is provided with a sample inlet 201 and a sample inlet 202, and the sample inlet 201 and the sample inlet 202 are specifically arranged on the second culture chamber 206. The culture may be added to the second culture chamber 206 through the sample addition port 201. And growth factors, medicines and the like can be added into the second culture cavity 206 to promote or inhibit the growth of the culture in the culture process according to experimental requirements, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into the system after the culture is completed to collect, treat or characterize the culture. A sample of the culture or broth may be taken for analysis during the culturing process via sampling port 202.

Specifically, the first exchange cavity 302 is provided with a first interface and a second interface, the liquid outlet is communicated with the first interface, and the premixing unit 1 is communicated with the second interface. The culture solution in the second culture cavity 206 can be conveyed into the first exchange cavity 302 through the liquid outlet and the first interface, and whether a power unit 5 is arranged between the first interface of the first exchange cavity 302 and the liquid outlet of the second culture cavity 206 can be determined according to the relative arrangement position of the first interface of the first exchange cavity 302 and the liquid outlet of the second culture cavity 206 and the conveying speed and the conveying amount of the culture solution, and the culture solution in the second culture cavity 206 is conveyed into the first exchange cavity 302 through the liquid outlet and the first interface by using the power unit 5. The culture solution in the first exchange cavity 302 can flow into the premixing cavity 101 through the second interface, and whether the power unit 5 is arranged between the second interface of the first exchange cavity 302 and the premixing cavity 101 or not can be determined according to the relative arrangement position of the second interface of the first exchange cavity 302 and the premixing cavity 101 and the requirements of the conveying speed and the conveying amount of the culture solution, and the culture solution in the first exchange cavity 302 is conveyed into the premixing cavity 101 through the second interface by using the power unit 5.

The fluid infusion unit 11 is in communication with the second exchange chamber 303, specifically, a third interface and a fourth interface are disposed on the second exchange chamber 303, and the third interface is in communication with the fluid infusion unit 11. The fluid replacement unit 11 is configured to deliver a supplemental nutrient solution to the second exchange chamber 303, and components in the supplemental nutrient solution in the second exchange chamber 303 can permeate through the membrane module into the first exchange chamber 302. A power unit 5 is arranged between the fluid infusion unit 11 and the second exchange cavity 303, and whether the power unit 5 is arranged between the fluid infusion unit 11 and the second exchange cavity 303 can be determined according to the relative arrangement position of the second exchange cavity 303 and the fluid infusion unit 11 and the requirements of the delivery speed and the delivery quantity of the supplementary nutrient fluid, and the supplementary nutrient fluid is delivered by using the power unit 5. The concentration of the supplementary nutrient solution is greater than that of the culture solution in the first exchange cavity 302, and the concentration difference between the two can drive components in the supplementary nutrient solution to move into the first exchange cavity 302 through the membrane component, and then the components are conveyed into the premixing cavity 101 to supplement the nutrient solution, so that the nutrient solution and the gas are mixed in the premixing cavity 101 to form the culture solution.

The collection unit 4 communicates with the second exchange chamber 303, and the collection unit 4 is used for collecting the culture solution discharged from the second exchange chamber 303. Specifically, the fourth interface communicates with the collection unit 4. The collection unit 4 communicates with the second exchange chamber 303 for collecting the culture liquid in the second exchange chamber 303. Preferably, in this embodiment, a power unit 5 is disposed between the collection unit 4 and the second exchange chamber 303, and the power unit 5 is used for conveying the culture solution in the second exchange chamber 303 into the collection unit 4. Depending on the relative arrangement of the second exchange chamber 303 and the collecting unit 4 and the requirements of the transport speed and transport amount of the culture liquid, it may be decided whether or not a power unit 5 is provided between the second exchange chamber 303 and the collecting unit 4, and the power unit 5 is used to transport the culture liquid in the second exchange chamber 303 to the collecting unit 4.

Example seven

As shown in fig. 14, the present embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, a fluid supplementing unit 11, a collecting unit 4 and a power unit 5.

At least one of the premixing unit 1, the culture chamber 2 of the culture unit, the fluid infusion unit 11 and the collection unit 4 is disposable, and is treated as waste after being used once, so that sanitary conditions in the perfusion culture process can be ensured, and normal perfusion culture is prevented from being influenced due to pollution of the premixing unit 1, the culture chamber 2, the fluid infusion unit and the collection unit 4.

The premix unit 1 comprises a premix chamber 101 and a gas premix control unit 105, wherein the culture medium contains nutrients required for the growth of the culture such as culture medium, factors, drugs and enzymes. The gas premix control unit 105 communicates with the premix chamber 101 for delivering gas [ including specifically oxygen, carbon dioxide, nitrogen, etc. ] into the premix chamber 101. The manner in which the premix chamber 101 may receive the nutrient solution, and the particular nutrient solution, will be described later and will not be described in detail herein. Gas is added to the premix chamber 101 by the gas premix control unit 105, and the introduced nutrient solution and gas are mixed in the premix chamber 101 to form a culture solution. The introduced oxygen can increase the dissolved oxygen amount of the culture solution, so that the dissolved oxygen amount can meet the growth requirement of the culture; the introduced carbon dioxide reduces and adjusts the pH value of the culture solution so as to meet the growth requirement of the culture. Preferably, in this embodiment, the number of the premixing chambers 101 and the number of the gas premixing control units 105 are one, and in other embodiments, the number of the premixing chambers 101 and the number of the gas premixing control units 105 are one, and the gas premixing control units 105 are simultaneously communicated with the plurality of premixing chambers 101, so that the gas concentration in the plurality of premixing chambers 101, in particular the dissolved oxygen concentration of the culture solution in the premixing chambers 101, can be controlled.

Preferably, the premixing unit 1 further comprises a pH detecting member 102, an oxygen dissolving detecting member 103 and a signal detector, wherein the pH detecting member 102 and the oxygen dissolving detecting member 103 are arranged in the premixing chamber 101, the signal detector can sense signals of the pH detecting member 102 and the oxygen dissolving detecting member 103, and the sensed signals are calculated and analyzed to obtain the pH value and the oxygen dissolving amount of the culture solution in the premixing chamber 101. With the change of the pH value and the dissolved oxygen amount in the premixing chamber 101, the signals detected by the pH detecting member 102 and the dissolved oxygen detecting member 103 will change, specifically, the optical signals of the pH detecting member 102 and the dissolved oxygen detecting member 103 will change, and the signal detector will periodically receive the optical signals of the surfaces of the pH detecting member 102 and the dissolved oxygen detecting member 103 to obtain data feedback, and the pH value and the dissolved oxygen amount are calculated according to the fed back data. The signal detector can feed back the detected information to the gas premix control unit 105, and if the dissolved oxygen is too low, the gas premix control unit 105 is caused to introduce oxygen into the premix chamber 101 to increase the dissolved oxygen of the culture fluid in the premix chamber 101. If the pH is too high, the gas premix control unit 105 is controlled to pass carbon dioxide to reduce the pH of the broth in the premix chamber 101. If the pH value is too low, lye is introduced into the premixing chamber 101 to increase the pH value of the culture solution in the premixing chamber 101.

The culture unit comprises a culture chamber 2, a first membrane assembly 203 is arranged in the culture chamber 2, and the first membrane assembly 203 divides the culture chamber 2 into a first culture cavity 205 and a second culture cavity 206. The first culture chamber 205 is used to hold a culture and to culture the culture. The culture chamber 2 is provided with a loading port 201 and a sampling port 202, and specifically, the loading port 201 and the sampling port 202 are both provided on the first culture chamber 205. The culture may be added to the first culture chamber 205 through the loading port 201. And growth factors, medicines and the like can be added into the first culture cavity 205 to promote or inhibit the growth of the culture in the culture process according to experimental requirements, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into the system after the culture is completed to collect, treat or characterize the culture. A sample of the culture or broth may be taken for analysis during the culturing process via sampling port 202.

The first culture chamber 205 is in communication with the premix chamber 101, and the premix chamber 101 is capable of delivering culture fluid to the first culture chamber 205. Preferably, in this embodiment, a power unit 5 is provided between the first culture chamber 205 and the premix chamber 101, and the power unit 5 is used to deliver culture fluid. The fluid infusion unit 11 is communicated with the second culture cavity 206, and the fluid infusion unit 11 is internally provided with a nutrient solution for providing the nutrient solution to the second culture cavity 206. Preferably, in the present embodiment, the power unit 5 is provided in the second culture chamber 206 and the fluid replacement unit 11, and the nutrient fluid is transported by using the power unit 5. The nutrient solution in the second culture chamber 206 can permeate through the first membrane module 203 into the first culture chamber 205. The concentration of the nutrient solution is greater than that of the culture solution in the first culture cavity 205, and the concentration difference between the nutrient solution and the culture solution can drive components in the nutrient solution to move into the first culture cavity 205 through the first membrane component 203 so as to supplement nutrient components required by the growth of the culture in the culture solution in the first culture cavity 205. Preferably, in this embodiment, the culture chambers 2 are one, i.e. one premix chamber 101 corresponds to one culture chamber. In other embodiments, the culture chambers 2 are multiple, and the culture chambers 2 are all communicated with the premixing chamber 101, i.e. the culture chambers 2 are connected in parallel, which can culture multiple cultures simultaneously, so that consistency of the cultures can be ensured in the subsequent test process, and accuracy of comparison of test results can be ensured.

Of the first culture chamber 205 and the second culture chamber 206, at least the second culture chamber 206 communicates with the collection unit 4, and preferably, waste generated by metabolism of the culture contained in the culture liquid in the first culture chamber 205 can permeate into the second culture chamber 206 through the first membrane module 203 due to a concentration difference, and thus, in this embodiment, both the first culture chamber 205 and the second culture chamber 206 communicate with the collection unit 4. A power unit 5 is provided between the first culture chamber 205 and the collection unit 4 and between the second culture chamber 206 and the collection unit 4, and the liquid is transported using the power unit 5. The nutrient solution in the second culture chamber 206 is discharged mainly when the concentration of metabolic waste of the culture in the second culture chamber 206 is too high or the concentration of the nutrient components required for the culture in the nutrient solution is too low, and then the new nutrient solution is replenished by using the replenishing unit 11.

The culture medium in the first culture chamber 205 and/or the second culture chamber 206 can be transferred into the premix chamber 101, and preferably the culture medium in the first culture chamber 205 can be transferred into the premix chamber 101 in this embodiment. Specifically, the first culture chamber 205 is provided with a liquid outlet, and the collecting unit 4 and the premixing chamber 101 are both communicated with the collecting unit 4, and more specifically, the in-vitro life support perfusion culture system further comprises a three-way valve 204, and three interfaces of the three-way valve 204 are respectively communicated with the liquid outlet, the collecting unit 4 and the premixing chamber 101 through pipelines. Preferably, in the present embodiment, a power unit 5 is provided between the three-way valve 204 and the drain port, and the culture medium is transported by using the power unit 5.

When the waste produced by the metabolism of the culture contained in the culture solution in the first culture chamber 205 is large, the three-way valve 204 is controlled to flow part of the culture solution in the first culture chamber 205 into the collecting unit 4; then, the three-way valve 204 is controlled to allow the culture solution in the first culture chamber 205 to flow into the premixing chamber 101, and the culture solution is mixed with the gas as needed in the premixing chamber 101 to form the culture solution, and then the culture solution is transferred into the first culture chamber 205 to replenish the culture solution in the first culture chamber 205.

Because the consumption speed of each component in the culture solution is different in the culture process, one or more components can be directionally added through the fluid infusion unit 11 according to the scheme of the embodiment, so that the balance of each component is ensured, the whole fluid exchange step is reduced, the cost is reduced, and the pollution probability of the culture is reduced. The fluid supplementing unit 11 supplements the second culture cavity 206 with fluid, and then the second culture cavity 206 conveys the nutrient fluid to the first culture cavity 205; therefore, the concentration and the composition of the nutrient solution in the fluid infusion unit 11 do not have to be formulated to be the same as those of the culture solution required for the culture, and the volume of the fluid infusion unit 11 can be made smaller. Meanwhile, the perfusion rate of the pre-mixing chamber 101 can be higher without worrying about excessively fast consumption of the culture solution due to the increase of the perfusion rate, improving the culture efficiency and increasing the kinds of cultures that can be cultured.

Specifically, the concentration of the culture medium component flowing out of the first culture chamber 205 can be detected and purposefully replenished by the replenishing unit 11. The concentration detection can be sampling detection or in-situ detection (such as infrared spectrum technology and fluorescence detection technology). Preferably, this embodiment employs fluorescence detection techniques. The system may also be provided with an automatic sample adding module for automatically adding nutritional components to the fluid replacement unit 11 based on the above concentration detection results.

detecting the concentration of a desired component of the culture in the culture solution in the first culture chamber 205 and/or determining the growth of the culture, and controlling the speed of the premix chamber 101 for feeding the culture solution into the first culture chamber 205 and the speed of the fluid replacement unit 11 for feeding the nutrient solution into the second culture chamber 206 according to the detected concentration and/or the determined growth of the culture.

Example eight

As shown in fig. 15, the present embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, a fluid supplementing unit 11, a collecting unit 4 and a power unit 5.

At least one of the premixing unit 1, the culture chamber 2 of the culture unit, the fluid infusion unit 11 and the collection unit 4 is disposable, and is treated as waste after being used once, so that sanitary conditions in the perfusion culture process can be ensured, and normal perfusion culture is prevented from being influenced due to pollution of the premixing unit 1, the culture chamber 2, the fluid infusion unit 11 and the collection unit 4.

The culture unit comprises a culture chamber 2, a first membrane assembly 203 is arranged in the culture chamber 2, and the first membrane assembly 203 divides the culture chamber 2 into a first culture cavity 205 and a second culture cavity 206. A third membrane module 207 is provided in the first culture chamber 205, and the third membrane module 207 divides the first culture chamber 205 into a first sub-culture chamber 2051 and a second sub-culture chamber 2052. The premix chamber 101 communicates with the first sub-culture chamber 2051, and the culture fluid in the premix chamber 101 can flow into the first sub-culture chamber 2051, and preferably a power unit 5 is provided in the first sub-culture chamber 2051 and the premix chamber 101, and the culture fluid is transported using the power unit 5. The specific arrangement form can be shown in fig. 16 or fig. 17. Preferably, in this embodiment, the culture chambers 2 are one, i.e. one premix chamber 101 corresponds to one culture chamber 2. In other embodiments, the culture chambers 2 are multiple, and the culture chambers 2 are all communicated with the premixing chamber 101, i.e. the culture chambers 2 are connected in parallel, which can culture multiple cultures simultaneously, so that consistency of the cultures can be ensured in the subsequent test process, and accuracy of comparison of test results can be ensured.

The second subculture chamber 2052 is used for placing a culture and culturing the culture. The culture chamber 2 is provided with a loading port 201 and a sampling port 202, and specifically, the loading port 201 and the sampling port 202 are provided on the second sub-culture chamber 2052. The culture may be added to the second subculture chamber 2052 through the sample addition port 201. And growth factors, medicines and the like can be added into the second sub-culture cavity 2052 to promote or inhibit the growth of the culture in the culture process according to experimental requirements, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into a system after the culture is completed to collect, treat or characterize the culture. A sample of the culture or broth may be taken for analysis during the culturing process via sampling port 202.

The third membrane module 207 is used to entrap and/or permeate components within the culture solution, and the third membrane module 207 is also capable of entrapping the culture, specifically entrapping cells of the culture, so that individual dispersed cells of the culture are not washed away, allowing the system to be used for more culture. Due to the consumption of the culture in the second sub-culture chamber 2052, the concentration of the components required for the culture in the culture solution in the second sub-culture chamber 2052 is lower than that in the first sub-culture chamber 2051, and the concentration of the waste produced by the metabolism of the culture is higher than that in the first sub-culture chamber 2051; useful components in the culture solution in the first subculture chamber 2051 permeate through the third membrane module 207 into the second subculture chamber 2052 for absorption and utilization of the culture, and metabolic waste in the second subculture chamber 2052 permeate through the third membrane module 207 into the first subculture chamber 2051.

First subculture chamber 2051 communicates with second culture chamber 206 through first membrane assembly 203. The fluid infusion unit 11 is communicated with the second culture cavity 206, and the fluid infusion unit 11 is internally provided with a nutrient solution for providing the nutrient solution to the second culture cavity 206. Preferably, in the present embodiment, the power unit 5 is provided in the second culture chamber 206 and the fluid replacement unit 11, and the nutrient fluid is transported by using the power unit 5. The nutrient solution in the second culture chamber 206 is able to permeate through the first membrane module 203 into the first culture chamber 205, specifically into the first subculture chamber 2051. The concentration of the nutrient solution is greater than that of the culture solution in the first subculture cavity 2051, and the concentration difference between the nutrient solution and the culture solution can drive components in the nutrient solution to move into the first subculture cavity 2051 through the first membrane assembly 203 so as to supplement nutrient components required by the growth of the culture in the culture solution in the first subculture cavity 2051.

Of the first culture chamber 205 and the second culture chamber 206, at least the second culture chamber 206 communicates with the collection unit 4. Due to the concentration difference, waste generated by the metabolism of the culture contained in the culture solution in the first culture chamber 205 can permeate into the second culture chamber 206 through the first membrane module 203, and preferably, the second culture chamber 206 is communicated with the collecting unit 4 in this embodiment, and waste generated by the metabolism of the culture in the second culture chamber 206 can be discharged into the collecting unit 4. The nutrient solution in the second culture chamber 206 is discharged mainly when the concentration of metabolic waste of the culture in the second culture chamber 206 is too high or the concentration of the nutrient components required for the culture in the nutrient solution is too low, and then the new nutrient solution is replenished by using the replenishing unit 11.

In other embodiments, it is also possible that both the first culture chamber 205 and the second culture chamber 206 are in communication with the collection unit 4, i.e. the first sub-culture chamber 2051 and/or the second sub-culture chamber 2052 are in communication with the collection unit 4, and the second culture chamber 206 is in communication with the collection unit 4. Specifically, as shown in FIG. 13, both the first sub culture chamber 2051 and the second culture chamber 206 are in communication with the collection unit 4. Due to the concentration difference, metabolic waste in the second sub-culture chamber 2052 permeates into the first sub-culture chamber 2051 through the third membrane module 207, and waste generated by metabolism of the culture contained in the culture solution in the first sub-culture chamber 2051 can permeate into the second culture chamber 206 through the first membrane module 203. Metabolites produced by the culture in first subculture chamber 2051 and second culture chamber 206 may be discharged into collection unit 4 and new liquid may be replenished in first subculture chamber 2051 and second culture chamber 206.

Preferably, the first subculture chamber 2051 and/or the second subculture chamber 2052 is capable of delivering culture medium to the premix unit 1, and in this embodiment, as shown, the culture medium in the first subculture chamber 2051 and the second culture chamber 206 is capable of being delivered into the premix chamber 101. The culture solution in the first sub-culture chamber 2051 and the second sub-culture chamber 2052 can be transported into the pre-mixing chamber 101 to be mixed with gas to supplement oxygen, and can be mixed with carbon dioxide or alkali liquor to adjust the pH value, so that the dissolved oxygen and the pH value of the culture solution flowing into the first sub-culture chamber 2051 from the pre-mixing chamber 101 meet the growth requirement of the culture.

In other embodiments, the culture solution in the second culture chamber 206 can be delivered into the premix chamber 101 to be converged into the premix chamber 101 for reuse; alternatively, the culture medium in the first sub-culture chamber 2051 can be fed into the premix chamber 101 to be pooled into the premix chamber 101 for reuse.

Optionally, the culture fluid in the second sub-culture chamber 2052 can be delivered into the first sub-culture chamber 2051, and in particular, the in vitro life support perfusion culture system further comprises a three-way valve 204, and three interfaces of the three-way valve 204 are respectively communicated with the first self-culture chamber 2051, the second sub-culture chamber 2052 and the premixing chamber 101 through pipelines. The culture medium in the second sub-culture chamber 2052 may be transferred to the first sub-culture chamber 2051 by adjustment of the three-way valve 204, or the culture medium in the second sub-culture chamber 2052 may be transferred to the premix chamber 101, or the culture medium in the first sub-culture chamber 2051 may be transferred to the premix chamber 101.

Example nine

As shown in fig. 18, the present embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, a fluid supplementing unit 11, a collecting unit 4 and a power unit 5.

The structure of the premix unit 1 is the same as in the eighth embodiment and will not be described in detail here.

The culture unit comprises a culture chamber 2, a first membrane assembly 203 is arranged in the culture chamber 2, and the first membrane assembly 203 divides the culture chamber 2 into a first culture cavity 205 and a second culture cavity 206. A third membrane module 207 is provided in the first culture chamber 205, and the third membrane module 207 divides the first culture chamber 205 into a first sub-culture chamber 2051 and a second sub-culture chamber 2052. The premix chamber 101 communicates with the second sub-culture chamber 2052, and the culture fluid in the premix chamber 101 can flow into the second sub-culture chamber 2052, and preferably a power unit 5 is provided in the second sub-culture chamber 2052 and the premix chamber 101, and the culture fluid is transported using the power unit 5. Preferably, in this embodiment, the culture chambers 2 are one, i.e. one premix chamber 101 corresponds to one culture chamber 2. In other embodiments, the culture chambers 2 are multiple, and the culture chambers 2 are all communicated with the premixing chamber 101, i.e. the culture chambers 2 are connected in parallel, which can culture multiple cultures simultaneously, so that consistency of the cultures can be ensured in the subsequent test process, and accuracy of comparison of test results can be ensured.

The first subculture chamber 2051 is used for placing a culture and culturing the culture. The culture chamber 2 is provided with a loading port 201 and a sampling port 202, and specifically, the loading port 201 and the sampling port 202 are provided on the first sub-culture chamber 2051. Culture may be added to first subculture chamber 2051 through sample addition port 201. And growth factors, medicines and the like can be added into the first sub-culture cavity 2051 to promote or inhibit the growth of the culture according to experimental requirements in the culture process, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into a system after the culture is completed to collect, treat or characterize the culture. A sample of the culture or broth may be taken for analysis during the culturing process via sampling port 202.

The third membrane module 207 is used to entrap and/or permeate components within the culture solution, and the third membrane module 207 is also capable of entrapping the culture, specifically entrapping cells of the culture, so that individual dispersed cells of the culture are not washed away, allowing the system to be used for more culture. Due to the consumption of the culture in the first subculture chamber 2051, the concentration of the components required for the culture in the culture medium in the first subculture chamber 2051 is lower than that in the second subculture chamber 2052, and the concentration of waste products generated by the metabolism of the culture is higher than that in the second subculture chamber 2052; useful components in the culture solution in the second sub-culture chamber 2052 permeate into the first sub-culture chamber 2051 through the third membrane module 207 for absorption and utilization of the culture, and metabolic waste in the first sub-culture chamber 2051 permeate into the second sub-culture chamber 2052 through the third membrane module 207.

Of the first culture chamber 205 and the second culture chamber 206, at least the second culture chamber 206 communicates with the collection unit 4. Due to the concentration difference, waste generated by the metabolism of the culture contained in the culture medium in the first sub-culture chamber 2051 can permeate into the second culture chamber 206 through the first membrane module 203, and preferably, the second culture chamber 206 is communicated with the collecting unit 4 in this embodiment, and waste generated by the metabolism of the culture in the second culture chamber 206 can be discharged into the collecting unit 4. The nutrient solution in the second culture chamber 206 is discharged mainly when the concentration of metabolic waste of the culture in the second culture chamber 206 is too high or the concentration of the nutrient components required for the culture in the nutrient solution is too low, and then the new nutrient solution is replenished by using the replenishing unit 11.

In other embodiments, it is also possible that both the first culture chamber 205 and the second culture chamber 206 are in communication with the collection unit 4, i.e. the first sub-culture chamber 2051 and/or the second sub-culture chamber 2052 are in communication with the collection unit 4, and the second culture chamber 206 is in communication with the collection unit 4. Specifically, both the first sub-culture chamber 2051 and the second culture chamber 206 are in communication with the collection unit 4. Waste products from the metabolism of the culture contained in the culture broth within first sub-culture chamber 2051 may permeate through first membrane assembly 203 into second culture chamber 206. Metabolites produced by the culture in first subculture chamber 2051 and second culture chamber 206 may be discharged into collection unit 4 and new liquid may be replenished in first subculture chamber 2051 and second culture chamber 206.

The first subculture chamber 2051 and/or the second subculture chamber 2052 is capable of delivering a culture solution to the premix unit 1 and preferably, in this embodiment, the culture solution in the first subculture chamber 2051 and the second culture chamber 206 is capable of being delivered into the premix chamber 101. The culture solution in the first sub-culture chamber 2051 and the second sub-culture chamber 2052 can be transported into the pre-mixing chamber 101 to be mixed with gas to supplement oxygen, and can be mixed with carbon dioxide or alkali liquor to adjust the pH value, so that the dissolved oxygen and the pH value of the culture solution flowing into the second sub-culture chamber 2052 from the pre-mixing chamber 101 meet the growth requirement of the culture.

In other embodiments, the culture solution in the second culture chamber 206 can be delivered into the premix chamber 101 to be converged into the premix chamber 101 for reuse; alternatively, as shown, the culture medium in the first sub-culture chamber 2051 can be transferred into the premix chamber 101 to allow the culture medium to be pooled into the premix chamber 101 for reuse.

Optionally, the culture fluid in the first sub-culture chamber 2051 can be transferred into the second sub-culture chamber 2052, and in particular, the in vitro life support perfusion culture system further comprises a three-way valve 204, and three interfaces of the three-way valve 204 are respectively communicated with the first self-culture chamber 2051, the second sub-culture chamber 2052 and the premixing chamber 101 through pipelines. The culture fluid in the first subculture chamber 2051 may be transferred to the second subculture chamber 2052 by adjustment of the three-way valve 204, or the culture fluid in the second subculture chamber 2052 may be transferred to the premix chamber 101, or the culture fluid in the first subculture chamber 2051 may be transferred to the premix chamber 101.

Examples ten

As shown in fig. 19, the present embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, a fluid supplementing unit 11, a collecting unit 4 and a power unit 5.

The culture unit comprises a culture chamber 2, a first membrane assembly 203 is arranged in the culture chamber 2, and the first membrane assembly 203 divides the culture chamber 2 into a first culture cavity 205 and a second culture cavity 206. A third membrane module 207 is provided in the first culture chamber 205, and the third membrane module 207 divides the first culture chamber 205 into a first sub-culture chamber 2051 and a second sub-culture chamber 2052. The premix chamber 101 communicates with the first sub-culture chamber 2051, and the culture fluid in the premix chamber 101 can flow into the first sub-culture chamber 2051, and preferably a power unit 5 is provided in the first sub-culture chamber 2051 and the premix chamber 101, and the culture fluid is transported using the power unit 5. Preferably, in this embodiment, the culture chambers 2 are one, i.e. one premix chamber 101 corresponds to one culture chamber 2. In other embodiments, the culture chambers 2 are multiple, and the culture chambers 2 are all communicated with the premixing chamber 101, i.e. the culture chambers 2 are connected in parallel, which can culture multiple cultures simultaneously, so that consistency of the cultures can be ensured in the subsequent test process, and accuracy of comparison of test results can be ensured.

First subculture chamber 2051 and second subculture chamber 2052 are each in communication with second culture chamber 206 through first membrane assembly 203. The fluid infusion unit 11 is communicated with the second culture cavity 206, and the fluid infusion unit 11 is internally provided with a nutrient solution for providing the nutrient solution to the second culture cavity 206. Preferably, in the present embodiment, the power unit 5 is provided in the second culture chamber 206 and the fluid replacement unit 11, and the nutrient fluid is transported by using the power unit 5. The nutrient solution in second culture chamber 206 is capable of penetrating into first culture chamber 205, specifically first subculture chamber 2051 and second subculture chamber 2052, through first membrane assembly 203. The concentration of the nutrient solution is greater than the concentration of the culture solution in the first subculture chamber 2051 and the second subculture chamber 2052, and the concentration difference between the two can drive the components in the nutrient solution to move into the first subculture chamber 2051 and the second subculture chamber 2052 through the first membrane assembly 203 so as to supplement the nutrient components required by the growth of the culture in the culture solution in the first subculture chamber 2051 and the second subculture chamber 2052.

Preferably, in this embodiment, the culture solution in the first sub-culture chamber 2051 can be conveyed into the premixing chamber 101, so that the culture solution in the first sub-culture chamber 2051 can be conveyed into the premixing chamber 101, and the first sub-culture chamber 2051 can be mixed with gas to supplement oxygen, and can be mixed with carbon dioxide or alkaline solution to adjust the pH value, so that the dissolved oxygen and the pH value of the culture solution flowing into the first sub-culture chamber 2051 from the premixing chamber 101 meet the growth requirement of the culture.

In other embodiments, the culture solution in the second culture chamber 206 can be delivered into the premix chamber 101 to be converged into the premix chamber 101 for reuse; alternatively, as shown, the culture medium in the first sub-culture chamber 2051 and the second culture chamber 206 can be transferred to the premix chamber 101 to allow the culture medium to be pooled into the premix chamber 101 for reuse.

It is to be understood that the above examples of the present invention are provided for clarity of illustration only and are not limiting of the embodiments of the present invention. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. Any modification, equivalent replacement, improvement, etc. which come within the spirit and principles of the invention are desired to be protected by the following claims.

Claims

1. An in vitro life support perfusion culture system, comprising:

a premix unit (1) comprising a premix chamber (101), adding a nutrient solution and a gas to the premix chamber (101), mixing within the premix chamber (101) to form a culture solution;

a culture unit comprising a culture chamber (2), wherein a first membrane component (203) is arranged in the culture chamber (2), the first membrane component (203) divides the culture chamber (2) into a first culture cavity (205) and a second culture cavity (206), and the first membrane component (203) is used for intercepting and/or penetrating partial components in culture solution;

the first culture cavity (205) is communicated with the premixing cavity (101), the premixing cavity (101) can convey culture solution to the first culture cavity (205), the culture solution in the first culture cavity (205) can flow back into the premixing cavity (101), and the second culture cavity (206) is used for containing culture and culturing the culture; or (b)

The first culture cavity (205) is communicated with the premixing cavity (101), the premixing cavity (101) can convey culture solution to the first culture cavity (205), the second culture cavity (206) is communicated with the premixing cavity (101), the culture solution in the second culture cavity (206) can flow back into the premixing cavity (101), and the first culture cavity (205) or the second culture cavity (206) is used for containing culture and culturing the culture;

A microscopic observation module (8) comprising a stage (801) on which the culture chambers (2) are placed, the stage (801) being capable of placing at least one of the culture chambers (2), and an observation assembly for observing the culture within the culture chambers (2);

the microscopic observation module (8) further comprises a rack (802), the objective table (801) is arranged on the rack (802) in a swinging manner, and the objective table (801) can drive the culture chamber (2) to swing synchronously;

the shaking amplitude and speed of the objective table (801) are matched with the speed of conveying the culture solution into the culture chamber (2); the lower culture solution conveying speed is matched with large-amplitude mechanical shaking, so that the culture solution can be updated simultaneously, the exchange opportunity of the nutrient solution and the culture is increased, and the nutrient substance exchange is promoted;

the first culture cavity (205) or the second culture cavity (206) is provided with a liquid outlet;

the in vitro life support perfusion culture system further comprises:

an exchange unit (3) comprising an exchange chamber and a second membrane assembly (301), wherein the exchange chamber comprises a first exchange cavity (302) and a second exchange cavity (303), the second membrane assembly (301) is arranged between the first exchange cavity (302) and the second exchange cavity (303), and is communicated with the first exchange cavity (302) through the second membrane assembly (301), and the liquid discharge port and the premixing unit (1) are both communicated with each other;

The second membrane component (301) entraps useful components in the culture solution in the first exchange cavity (302), and metabolites generated by the growth of the culture in the first exchange cavity (302) permeate into the second exchange cavity (303) through the second membrane component (301) and are discharged from the second exchange cavity (303).

2. The in vitro life support perfusion culture system according to claim 1, wherein the pre-mix unit (1) further comprises a fluid replacement chamber (104) in communication with the pre-mix chamber (101) for delivering a nutrient fluid into the pre-mix chamber (101), the fluid replacement chamber (104) being capable of directionally delivering one or more components required for the culture into the pre-mix chamber (101).

3. The in vitro life support perfusion culture system of claim 1, wherein the in vitro life support perfusion culture system comprises,

the first culture cavity (205) is provided with a liquid inlet which is communicated with the premixing chamber (101);

the in vitro life support perfusion culture system further comprises:

and the fluid supplementing unit (11) is communicated with the second exchange cavity (303) and is used for conveying the supplementary nutrient fluid to the second exchange cavity (303), the fluid supplementing unit (11) can directionally convey one or more components required by the culture into the second exchange cavity (303), and the components in the supplementary nutrient fluid can permeate into the first exchange cavity (302) through the second membrane assembly (301).

4. An in vitro life support perfusion culture system according to claim 3, further comprising a collection unit (4), the collection unit (4) being in communication with the second exchange chamber (303), the collection unit (4) being adapted to collect the culture fluid discharged from the second exchange chamber (303).

5. The in vitro life support perfusion culture system according to claim 4, further comprising a power unit (5), wherein the power unit (5) is arranged between the collection unit (4) and the second exchange chamber (303), and/or wherein the power unit (5) is arranged between the fluid infusion unit (11) and the second exchange chamber (303).

6. The in vitro life support perfusion culture system according to claim 1, wherein the pre-mixing unit (1) further comprises a pH detector (102), a dissolved oxygen detector (103) and a signal detector, wherein the pH detector (102) and the dissolved oxygen detector (103) are both arranged in the pre-mixing chamber (101), and the signal detector can sense signals fed back by the pH detector (102) and the dissolved oxygen detector (103) so as to obtain the pH value and the dissolved oxygen amount of the culture solution in the pre-mixing chamber (101).

7. The in vitro life support perfusion culture system according to claim 6, wherein the pH detection member (102) is a pH electrode plate, and the dissolved oxygen detection member (103) is a dissolved oxygen electrode plate;

The pH electrode plate is arranged on the inner wall of the premixing chamber (101) or immersed in the liquid of the premixing chamber (101), and the dissolved oxygen detector (103) is arranged on the inner wall of the premixing chamber (101) or immersed in the liquid of the premixing chamber (101).

8. The in vitro life support perfusion culture system according to claim 1, wherein the pre-mixing unit (1) further comprises a gas pre-mixing control unit (105) in communication with the pre-mixing chamber (101) for delivering gas into the pre-mixing chamber (101).

9. The in vitro life support perfusion culture system according to claim 4, wherein at least one of the pre-mix unit (1), the culture chamber (2) and the collection unit (4) is a disposable consumable.

10. The in vitro life support perfusion culture system according to any of claims 1 to 9, wherein the culture chamber (2) is a plurality, wherein a plurality of culture chambers (2) are each in communication with the premix chamber (101).

11. The in vitro life support perfusion culture system of any one of claims 1-9, further comprising:

the temperature control unit (6) comprises a refrigeration assembly and a temperature control module, wherein the refrigeration assembly is electrically connected with the temperature control module, and the temperature control module is used for controlling the refrigeration assembly to cool the culture chamber (2) to a first preset temperature, and the first preset temperature is the liquefaction temperature of the culture supporting structure;

A sampling port (202) is arranged on the culture chamber (2), and the sampling port (202) is used for taking out the culture after the culture supporting structure is liquefied.

12. The in vitro life support perfusion culture system of any one of claims 1-9, further comprising:

the sterile control module (9) comprises a sterile working chamber (901), a filtering assembly (903) and a sterilizing assembly (904), wherein at least the culture chamber (2) is arranged in the sterile working chamber (901), the filtering assembly (903) is used for filtering gas which is introduced into the sterile working chamber (901), and the sterilizing assembly (904) is used for sterilizing the sterile working chamber (901).

13. The in vitro life support perfusion culture system according to any one of claims 1 to 9, wherein the culture unit further comprises a mixing module (7) for shaking the culture fluid inside the culture chamber (2).

14. The in vitro life support perfusion culture system according to claim 13, wherein the number of premixing chambers (101) is a plurality, the number of gas premixing control units (105) is a single, and the gas premixing control units are simultaneously communicated with the plurality of premixing chambers (101) and can control the gas concentration in the plurality of premixing chambers (101).

15. A method of controlling an in vitro life support perfusion culture system according to any one of claims 1-9, comprising the steps of:

detecting the concentration of a desired component of the culture in the culture chamber (2) and/or determining the growth of the culture, and controlling the speed of the transfer of the culture fluid from the premix chamber (101) to the culture chamber (2) based on the detected concentration and/or the determined growth of the culture.