CN109337813B

CN109337813B - System and method suitable for biological tissue culture and real-time monitoring

Info

Publication number: CN109337813B
Application number: CN201811226234.2A
Authority: CN
Inventors: 王玲; 吕晨泽; 徐铭恩; 斯培剑; 戴嘉韵; 赖雪聪
Original assignee: Regenovo Biotechnology Co ltd
Current assignee: Regenovo Biotechnology Co ltd
Priority date: 2018-10-19
Filing date: 2018-10-19
Publication date: 2023-07-11
Anticipated expiration: 2038-10-19
Also published as: CN109337813A

Abstract

The invention provides a system and a method suitable for biological tissue culture and real-time monitoring, which relate to the technical field of tissue engineering and comprise the following steps: the biological 3D printer, the organ chip, the connecting base, the driving system and the auxiliary system; the organ chip is connected with the driving system through the connecting base; the biological 3D printer is used for constructing biological 3D printing tissue; the organ chip is used for placing a culture medium and biological 3D printing tissues and culturing the biological 3D printing tissues; the connecting base is used for placing an organ chip and is connected with the driving system, the driving system is used for driving the culture medium to flow in the organ chip, and the auxiliary system is used for monitoring the state of biological 3D printing tissue. The system and the method provide a novel culture platform for tissue culture, pathology research, drug screening and other works.

Description

System and method suitable for biological tissue culture and real-time monitoring

Technical Field

The invention relates to the field of tissue engineering, in particular to a system and a method suitable for biological tissue culture and real-time monitoring.

Background

The organ chip provides a novel culture platform for cell or tissue culture, pathology research, drug screening and other works, and has the main characteristic of being capable of more effectively simulating interaction of organs in a human body. Organ-chip involves three key elements, living tissue/organ elements, fluid control elements, detection/sensing elements, respectively. The fluid control element provides living cell tissue living substrate and continuous perfusion culture mode to simulate in-vivo growth microenvironment; living tissue/organ elements refer to components that spatially align a particular cell type in 2D or 3D; the detection/sensing element provides monitoring and evaluation functions.

Although 3D tissue structures can better mimic in vivo conditions compared to 2D conditions, the consistency of 3D tissue preparation, the controllability of three-dimensional culture, and the real-time monitorability of 3D cell lines are all difficulties, and how to prepare standardized 3D living tissue, construct three-dimensional culture systems, and three-dimensional monitoring schemes remain challenges. Biological 3D printing provides a highly consistent and controllable organoid tissue culture method, but in the existing organ-chip technology, 3D tissue cultured organoids are used more because of the inconvenience of directly printing biological 3D printed organoids on the organ chip.

Disclosure of Invention

In view of the above, the present invention aims to provide a system and a method suitable for biological tissue culture and real-time monitoring, which solve the problem of organic combination of organ chip 3D tissue construction, perfusion culture and monitoring.

In a first aspect, an embodiment of the present invention provides a system suitable for biological tissue culture and real-time monitoring, including: the biological 3D printer, the organ chip, the connecting base, the driving system and the auxiliary system; the organ chip is connected with the driving system through a connecting base;

the biological 3D printer is used for constructing biological 3D printing tissue;

The organ chip is used for placing a culture medium and biological 3D printing tissues and culturing the biological 3D printing tissues;

the connecting base is used for placing the organ chip and is connected with the driving system;

the driving system is used for driving the culture medium to flow in the organ chip;

the auxiliary system is used for monitoring the state of biological 3D printing tissue.

With reference to the first aspect, the embodiment of the present invention provides a first possible implementation manner of the first aspect, wherein the organ-chip includes a transparent top cover, a transport unit, and an organ-chip main body; the transfer unit is detachably embedded into the organ chip main body, the transparent top cover covers the organ chip main body, and the transfer unit is used for containing biological 3D printing tissues.

With reference to the first aspect, the embodiment of the present invention provides a second possible implementation manner of the first aspect, wherein the organ-chip main body further includes: the micro-channel layer is arranged between the hard top layer and the transparent bottom layer, and the sensing chip is in contact with a culture medium of the organ chip main body;

The hard top layer comprises at least one culture chamber, a gas channel, a top surface groove, a bottom surface groove and a detection area; the bottom surface groove is used for placing the micro-channel layer;

the micro-channel layer comprises at least one culture chamber, a micro-channel, a driving groove, a liquid storage groove, a dividing groove and a fence-shaped valve; the micro-flow channel connects the at least one culture chamber, the driving groove, the liquid storage groove and the dividing groove; the fence-shaped valve separates and breaks the dividing grooves;

wherein the transfer unit is matched with at least one culture chamber in the hard top layer and at least one culture chamber in the micro-channel layer.

With reference to the first aspect, the embodiment of the present invention provides a third possible implementation manner of the first aspect, wherein the organ-chip main body is rectangular or hexahedral in shape.

With reference to the first aspect, an embodiment of the present invention provides a fourth possible implementation manner of the first aspect, wherein the auxiliary system includes: an observation system, and/or an analysis system, and/or a control system, and/or an environmental control system;

the observation system is used for observing the cultured test tissues and the culture medium;

The analysis system is used for analyzing according to the sensing chip to obtain detection data;

the control system is used for accurately controlling the driving system to control the flow speed and the flow direction of the culture medium in the micro-channel;

the environmental control system provides suitable temperature, humidity and carbon dioxide concentration for biological 3D printing tissue growth.

With reference to the first aspect, an embodiment of the present invention provides a fifth possible implementation manner of the first aspect, wherein the driving system includes a display control device, a control device, and an air path distribution device; the control device is respectively connected with the display control device and the air path distribution device;

the display control device is used for displaying a plurality of working modes so that a user can select according to the plurality of working modes displayed in a viewing mode;

the control device is used for controlling the air path distribution device to work according to the working mode selected by the user.

With reference to the first aspect, an embodiment of the present invention provides a sixth possible implementation manner of the first aspect, where the system further includes a connection base, where the connection base is used in combination with the driving system by a connection manner or is used in combination with the pneumatic system as a whole.

With reference to the first aspect, an embodiment of the present invention provides a seventh possible implementation manner of the first aspect, where the system further includes: the driving system is connected with the organ chip through the updating system and is also used for driving the updating system to update the culture medium in the organ chip.

In a second aspect, an embodiment of the present invention further provides a method suitable for biological tissue culture and real-time monitoring, including:

using a biological 3D printer to construct a biological 3D printed tissue for testing/testing; the 3D printing tissue is used as a test tissue to be directly printed in the organ chip culture room, or is printed on the transfer unit and then placed in the organ chip culture room;

culturing the test tissue in the organ-chip according to the working mode selected by the user by receiving the working mode selected by the user;

and in the culture process, detecting the test tissue to obtain detection data.

With reference to the second aspect, an embodiment of the present invention provides a first possible implementation manner of the second aspect, wherein the culturing the test tissue in the organ-chip according to the operation mode selected by the user by receiving the operation mode selected by the user includes:

After receiving the working mode selected by the user, the positive pressure air source and the negative pressure air source are controlled to output positive pressure air flow and negative pressure air flow to the air channel distribution device, so that the air channel distribution device leads the positive pressure air flow and the negative pressure air flow to the air channel of the organ chip to enable the culture medium to move.

With reference to the second aspect, an embodiment of the present invention provides a second possible implementation manner of the second aspect, wherein, by receiving a user-selected operation mode, after the step of culturing the test tissue in the organ-chip according to the user-selected operation mode, the method further includes:

when the culture medium in the organ chip is determined to be stored in the bubble, the current working mode is adjusted according to the position of the bubble.

The embodiment of the invention has the following beneficial effects: by adding a biological 3D printer in the system, in the test, the biological 3D printer is used for constructing biological 3D printing tissue for test/test, a culture medium and the biological 3D printing tissue are placed on an organ chip, the biological 3D printing tissue is cultured, a driving system drives the culture medium to flow in the organ chip, and an auxiliary system monitors the biological 3D printing tissue and the culture medium.

Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.

In order to make the above objects, features and advantages of the present invention more comprehensible, preferred embodiments accompanied with figures are described in detail below.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.

FIG. 1a is a block diagram of a system suitable for biological tissue culture and real-time monitoring according to one embodiment of the present invention;

FIG. 1b is a block diagram of a system suitable for biological tissue culture and real-time monitoring according to another embodiment of the present invention;

FIG. 2 is a block diagram of an organ chip according to an embodiment of the invention;

fig. 3a is a schematic diagram of the front surface of a micro flow channel layer according to an embodiment of the present invention;

FIG. 3b is a schematic diagram of the reverse side of the micro flow channel layer according to the embodiment of the present invention;

FIG. 4 is a diagram showing a structure of a medium replacement method in an organ-chip according to an embodiment of the present invention;

FIG. 5 is a block diagram of a drive system according to an embodiment of the present invention;

FIG. 6 is a graph showing a gas circuit distribution diagram according to an embodiment of the present invention;

FIG. 7 is a block diagram of a connection base according to an embodiment of the present invention;

FIG. 8 is a block diagram of a ventilation module according to an embodiment of the present invention;

fig. 9 is a flowchart of a method for biological tissue culture and real-time monitoring according to an embodiment of the present invention.

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 present invention will be clearly and completely described below with reference to the accompanying drawings, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. 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.

The organ tissue of the existing organ chip grows on the tissue culture module of the chip in an adherence way, and the design can increase the death rate of cells or cause the blockage of a micro flow channel due to the massive proliferation and shedding of cells.

The existing organ chip adopts a tissue culture module with 2D adherence growing on the chip, the death rate of cells is increased or a micro flow channel is blocked due to the large amount of proliferation and shedding of cells, the tissue structure of the design is far from the in-vivo real situation, the 3D tissue structure can better simulate the in-vivo situation, but the consistency of 3D tissue preparation, the controllability of three-dimensional culture and the real-time monitoring have difficulties compared with the 2D cell tissue culture module, and the preparation of standardized 3D living tissue, the construction of a three-dimensional culture system and a three-dimensional monitoring scheme are still challenges.

Based on the above, the system and the method for biological tissue culture and real-time monitoring provided by the embodiments of the present invention provide a novel culture platform for tissue culture, pathology research, drug screening and other works, and are mainly characterized by providing a tissue-like organ which is closer to a real tissue, more effectively simulating interaction of organs in a human body, and providing a real-time continuous monitoring method, by adding a biological 3D printer in the system, in the test, the biological 3D printer is configured to test/test biological 3D printing tissue, an organ chip is configured to place a culture medium and the biological 3D printing tissue, the biological 3D printing tissue is cultured, a driving system drives the culture medium to flow in the organ chip, and the auxiliary system monitors the biological 3D printing tissue and the culture medium.

For the convenience of understanding the present embodiment, a system and method for biological tissue culture and real-time monitoring disclosed in the present embodiment will be described in detail.

Referring to FIG. 1a, the present invention provides a system suitable for biological tissue culture and real-time monitoring, comprising: biological 3D printer 1000, organ chip 2000, drive system 3000, auxiliary system 4000, and connection base 5000; organ chip 2000 is connected to drive system 3000 via connection base 5000, and biological 3D printer 1000, organ chip 2000, drive system 3000, auxiliary system 4000, and connection base 5000 may all be provided on the same laboratory bench.

The biological 3D printer 1000 is used to construct biological 3D printed tissue for testing/testing, among other things. Organ chip 2000 is used for placing culture medium and biological 3D printing class tissue, cultivates biological 3D printing class tissue. The drive system 3000 is used to drive the flow of the culture medium within the organ-chip. The auxiliary system 4000 is used to monitor biological 3D printing tissue and/or culture medium.

Specifically, when biological 3D printing tissue is printed out using biological 3D printer 1000 and then placed in organ chip 2000, biological 3D printing tissue and culture medium are present in organ chip 2000, and the driving system drives the culture medium to flow in organ chip 2000, thereby culturing biological 3D printing tissue. The auxiliary system can detect biological 3D printing tissue and/or culture medium to obtain detection data.

For example, the assist system 4000 can detect cell proliferation in test tissue, or cell growth status in test tissue, or biological slicing results in biological 3D-printed tissue, and so forth.

Types of biological 3D printing-like tissues may include, but are not limited to: liver, kidney, skin, small intestine, lung, tumor and heart.

Organ-chip 2000 also includes a transparent top cover, a transfer unit, and an organ-chip body; the transport unit is detachably embedded into the organ chip main body, the transparent top cover covers the organ chip main body, and the transport unit is used for containing the biological 3D printing tissue.

Referring to fig. 2, the transfer unit 2100 is embedded inside an organ-chip main body including: the micro flow channel layer 2300 is disposed between the hard top layer 2200 and the transparent bottom layer 2400, the micro flow channel layer 2300 is in contact with the culture medium of the organ-chip body, and the sensing chip may be disposed between the micro flow channel layer 2300 and the transparent bottom layer 2400.

Wherein the rigid top layer 2200 is made of a transparent, non-toxic material to cells, which may include, but is not limited to, polystyrene Plastic (PS), polycarbonate (PC), cyclic olefin copolymer plastic (COC plastic), polyethylene (PE), polypropylene (PP).

The rigid top layer 2200 includes at least one culture chamber, a top surface recess, a bottom surface recess, and a detection area 2210; the bottom surface groove is used for placing the micro-channel layer.

At least one culture room in the hard top layer can be used for placing test products such as cells, tissues or biological 3D printing tissues and the like for test, and can also be used for placing transport units containing the cells, the tissues or the biological 3D printing tissues; and the culture room comprises positioning points and positioning eaves, and can be matched with the positioning openings of the transfer units.

The grooves on the top surface can distinguish different micro-channel layers and are matched with the serial numbers of the culture chambers for use. Detection area 2210 can be used for placing 96-well plate ELISA strips, analyzing and detecting the extracted culture medium, or directly detecting in the detection area. The transparent top cover clamping groove is used for placing the transparent top cover;

as described with reference to fig. 3a and 3b, the micro flow channel layer 2300 includes at least one culture chamber 2310, a micro flow channel 2320, a check valve, a driving groove 2330, a liquid storage groove 2340, a dividing groove 2350, and a barrier-shaped valve 2360; the micro flow channel 2320 connects the at least one culture chamber 2310, the one-way valve, the driving groove 2330, the liquid storage groove 2340 and the dividing groove 2350; the barrier valve 2360 breaks the dividing groove partition 2350.

The micro flow channel layer 2300 is composed of Polydimethylsiloxane (PDMS) or a soft and cell growth-harmless material containing polydimethylsiloxane, and both sides thereof are bonded to the hard top layer 2200 and the transparent bottom layer 2400 by oxygen plasma treatment in an irreversible bonding manner.

The micro flow channel 2320 is connected with the culture chamber 2310 in the micro flow channel layer, the flow channel width gradient, the dividing groove 2350 and the liquid storage groove 2340 are separated in the dividing groove 2350 by the fence-shaped valve 2360, or a one-way valve can be added in the micro flow channel 2320 to control the flow direction and prevent backflow; the diameter of the micro flow channel is less than or equal to 1mm, a circulating flowing liquid culture medium is provided for a culture chamber of the micro flow channel layer, nutrition is provided for the growth of cells, tissues or biological 3D printing tissues, and metabolic waste is discharged or the culture medium containing the metabolic waste is discharged into the next culture chamber;

alternatively, the micro flow channel 2320 has a section of micro flow channel with gradually changed width, which may be referred to as the flow channel width gradient, where the width of the flow channel width gradient near one end of the culture chamber is greater than the width near one end of the micro flow channel, or the flow channel width gradient is a section of micro flow channel with gradually changed diameter when the micro flow channel and the culture chamber are connected through a vertical flow channel.

Further, the driving system 3000 operates according to the following principle: since the driving system 3000 uses the difference of air pressure to control the flow of the culture medium in the organ chip 2000 and the culture medium is mainly in the hard top layer 2200 and the micro channel layer 2300, the air passage of the hard top layer 2200 is described first. In the invention, the hard top layer 2200 comprises a gas channel, the gas channel comprises a gas pipe interface and a gas output port, the gas pipe interface and the gas output port can be communicated together through a conduit to form a gas channel, and the gas output port can be connected with the driving groove 2330 of the micro-channel layer 2300; the air pipe interface can be connected with the driving system 3000 through a pipeline, and the air output port on the hard top layer corresponds to the position and the shape of the driving groove in the micro-channel layer. In the micro flow channel layer 3200, a driving groove may be separated from the dividing groove 2350 or the liquid storage groove 2340 using a flexible film; the driving groove 2330 corresponds to the position and the shape of the gas outlet on the hard top layer, and can deform the soft film by changing the air pressure in the air channel, so that the volumes of the dividing groove 2350 and the liquid storage groove 2340 are changed, the opening and closing of the fence-shaped valve 2360 and the control of the liquid volume in the liquid storage groove 2340 are realized, and the flow of the culture medium in the organ chip is realized.

In the case of internal circulation of the culture medium in the organ-chip main body, the micro flow channel layer may have the following structure, and referring to fig. 3a, the micro flow channel 3220 may be a closed loop flow channel, and the culture medium in the micro flow channel is self-circulated by the driving system 3000, and the update of the culture medium is realized by periodically replacing the culture medium manually. The manual periodic replacement can be performed by taking out the original culture medium and then placing the new culture medium into the organ-chip main body by using a tool capable of containing liquid.

Or the microchannels may be made to circulate in one or more height dimensions, i.e. the culture medium may flow in the same plane of the channels, or may enter the channels of another height dimension through a vertical channel.

When the driving system is used to update the culture medium in the organ-chip main body, as shown in fig. 4, the driving system may be connected to the organ-chip 2000 through the updating system, where the updating system includes a liquid storage tank 6100 and a waste liquid tank 6200, the driving system is further used to drive the updating system to update the culture medium in the organ-chip, specifically, the driving system 3000 is connected to the liquid storage tank 6100 through a pipeline, the liquid storage tank 6100 is connected to the organ-chip 2000 through a pipeline, the waste liquid tank 6200 is connected to the organ-chip 2000 through a pipeline, and the surface of the hard top layer 2200 may be provided with a liquid inlet 2220 and a liquid outlet 2230 as required. Specifically, the new culture medium is driven by the drive system to enter liquid inlet 2220 through the pipeline, and after circulation in organ-chip 2000 is completed, the new culture medium leaves organ-chip 2000 through liquid outlet 2230 through the pipeline.

Wherein, the hard top layer 2200, the micro flow channel layer 2300 and the transparent bottom layer 2400 are bonded by an oxygen plasma treatment in an irreversible bonding manner, and the culture chamber of the hard top layer 2200 is correspondingly matched with the culture chamber 2310 of the micro flow channel layer 2300 during the bonding process, and the micro flow channel layer 2300 and the transparent bottom layer 2400 fix the sensing chip between the micro flow channel layer 2300 and the transparent bottom layer 2400 during the bonding process, and can be contacted with the culture medium in the micro flow channel 2320 of the micro flow channel layer 2300, and the organ chip body is used as a whole.

The transparent bottom layer 2400 may be composed of a transparent hard glass or other transparent hard material, irreversibly bonded to the micro flow channel layer 2300, provide a substrate for the micro flow channel layer 2300, and contain a sensor chip.

The sensing chip is a biochip with a surface functionalized with a biomarker, is arranged at the top end or the bottom end of a culture chamber in a micro-channel, a hard top layer or a micro-channel layer and is contacted with a culture medium; according to the selected sensing principle and the sensing conditions required by the sensing chip, a channel for leading the sensing conditions to enter and exit the organ chip is arranged on the organ chip main body, and the auxiliary system is communicated with the sensing chip; the sensed conditions include, but are not limited to, light, current, voltage, magnetic field, and various aspects of the organ-chip characteristics are detected and recorded in real-time.

In addition, the biological 3D printer can print directly in the culture room on the hard top layer; or the biological 3D printing tissue is printed in a transfer unit and then placed in a culture room on the organ chip main body.

Referring to fig. 5, the driving system 3000 includes a display control device 3100, a control device 3200, and a gas path distribution device 3300;

the display control device 3100 may include a touch screen 3130, an indicator light 3110, and a switch 3120, and is in information connection with the control device, and is configured to display a plurality of operation modes, and a user may select an operation mode desired by the user by viewing the operation mode on the touch screen 3130. The indicator light 3110 may be used to indicate that the touch screen may be used, and the switch 3120 may be used to activate the display control device.

The control device is used for controlling the air path distribution device to work according to the working mode selected by the user. Specifically, as shown in fig. 5, the control device 3200 outputs positive pressure and negative pressure air flows to the air path distribution device 3300 by controlling the positive pressure air source and the negative pressure air source, so that the air path distribution device 3300 leads the positive pressure and the negative pressure to the air channel of the organ-chip main body. The gas path distribution device 3300 is connected with the organ-chip main body through a quick plug and a gas pipe, and is connected with at least one organ-chip main body; the air path distribution device controls positive pressure and negative pressure of air flow output to the organ chip main body, the culture medium flows clockwise or anticlockwise through a circulating action formed by four steps, when the culture medium flows clockwise, the circulation is clockwise, and when the culture medium flows anticlockwise, the circulation is anticlockwise. The circulation formed by the four steps is completed by controlling positive pressure or negative pressure output of the three driving grooves;

A first dividing groove, a liquid storage groove and a second dividing groove are arranged below the soft film corresponding to the three driving grooves; in operation, as shown in fig. 6, the air pressures corresponding to the three driving grooves are respectively: step 1, outputting positive pressure, negative pressure and negative pressure; step 2, outputting negative pressure, positive pressure and negative pressure; step 3, outputting negative pressure, positive pressure and positive pressure; step 4, outputting negative pressure, negative pressure and positive pressure; the driving of the culture medium from the first dividing groove to the second dividing groove is completed.

Wherein the working mode selection comprises a liquid adding mode, an alternating mode, a circulating mode, a manual mode and a stepping mode;

the liquid adding mode is that all gas channels provide negative pressure, so that all soft films of the micro-channel layer are lifted, all fence-shaped valves are opened, the volume of a liquid storage groove is increased, and a culture medium is added for an organ chip.

The alternating mode means that the gas path distribution device can be set through parameter setting, so that clockwise or anticlockwise circulation alternation is realized. During actual control, the control device outputs positive pressure and/or negative pressure to the gas path distribution device, and the gas path distribution device outputs gas paths, so that the culture medium flows for a period of time according to clockwise, flows for a period of time according to anticlockwise and circularly and alternately.

The circulation mode is to set the gas path distribution device to continuously perform clockwise or anticlockwise circulation through parameter setting. In actual control, the control device outputs positive pressure and/or negative pressure to the gas path distribution device, and the gas path distribution device outputs gas paths so that the culture medium flows clockwise or anticlockwise.

The manual mode is to manually control the positive pressure output and the negative pressure output of all the air path channels through the display control device.

The step mode refers to the last step or the next step of the set anticlockwise or clockwise cycle of adjusting the state of the air channel through the display control device. For example, when the flow is clockwise, in the step mode, the state of the gas path channel can be operated to enter the next clockwise circulation step, and one clockwise circulation is completed through four operations; or when the air flows clockwise, the state of the air channel enters the previous clockwise circulation step through operation, and one anticlockwise circulation is completed through four operations.

Wherein, auxiliary system 4000 includes: an observation system and/or an analysis system. The observation system is used for observing the test tissue and the culture medium; the analysis system is used for analyzing according to the sensing chip to obtain detection data.

In addition, the auxiliary system also comprises a control system and an environment control system.

The control system precisely controls the driving system 3000 to control the flow speed and the flow direction of the culture medium in the micro-channel; the reaction of the biological 3D printing tissue in the organ chip main body culture room and substances in the culture solution is realized by controlling the exchange time of the flowing culture medium.

The observation system utilizes the high transparency of the organ chip to complete the function of directly observing the biological 3D printing tissue cultured in the culture room and the culture medium in the micro-channel.

Wherein the observation system may select, but is not limited to, the following methods: optical microscopy, fluorescence microscopy, phase contrast microscopy, optical Coherence Tomography (OCT);

the environmental control system provides the proper temperature, humidity and carbon dioxide concentration for tissue growth. The environment control system mainly provides environment factors such as temperature, humidity, carbon dioxide concentration and the like for proper tissue culture in the incubator.

In addition, the pH, temperature, flow rate and other information can be sent to the environment control system by connecting a sensing chip capable of monitoring the indexes such as the pH, the temperature, the flow rate and the like in the micro-channel, the environment control system can monitor the pH, the temperature, the flow rate and other information according to the acquired information, and when the condition of the excess value is found, a user can be reminded through an alarm.

The analysis system can analyze the signals obtained from the sensing chip and provide an indirect real-time online monitoring for the biological 3D printing tissue cultured on the organ chip through the analysis result;

when the signal output by the sensing chip is an optical signal, the analysis system needs to have the detection capability of one or more of parameters such as wavelength, power, energy, incident angle, pulse width, repetition frequency, polarization degree and the like of light;

when the signal output by the sensing chip is an electrical signal, the analysis system needs to have the capability of detecting one or more parameters of voltage, current, charge quantity and the like of the circuit.

And in conjunction with fig. 1, connection mount 5000 is used in conjunction with organ chip 2000. The connecting base can be used in combination with the driving system in a connecting mode, or the connecting base and the driving system are combined into a whole for use. Referring again to fig. 1, drive system 3000 is coupled to a connection mount via conduit 3500.

Referring to fig. 7, the connection base 5000 includes a base cover 5010, a fixing bayonet 5020, a clamping groove 5030, a ventilation module 5040, a ventilation slider 5041, an air guide connector 5042, a spring post, a quick connector 5044, an air guide pipe 5045, a sliding group 5050, an observation window, and a rotating shaft 5070;

As shown in fig. 8, the air duct of the ventilation module 5040 is connected to the quick connector 5044, the air duct connector 5042 and the quick connector 5044 are connected to two sides of the ventilation slider 5041, and are mutually communicated through the air duct inside the ventilation slider; the ventilation module 5040 is connected with the clamping groove 5030 through the sliding group 5050, and the sliding group 5050 and the ventilation module 5040 can slide relative to the clamping groove.

The base cover 5010 is connected with the clamping groove 5030 through the rotating shaft 5070, and the base cover 5010 can push the ventilation module 5040 to slide forwards in the covering process; the base cover is opened to more than 90 degrees, the air pipe interface surface of the organ chip is placed into the clamping groove 5030 connected with the base in the direction of the air guide connector 5042, the base cover 5010 is pressed downwards by hands, the air guide module 5040 is pushed to slide forwards by the base cover 5010 in the covering process, the air guide connector 5042 is inserted into the air pipe interface of the organ chip body, and the air guide connector 5042 is sealed through the O-shaped sealing ring on the air guide connector 5042. After the base cover 5010 is covered, the fixing bayonet 5020 is adjusted to fix the base cover 5010, so that looseness is prevented.

In addition to the form of fig. 1a, the present invention also provides an organ chip having the shape described in connection with fig. 1b, which may be made as a hexahedron, and the organ chip 2000 includes a transparent top cover 2500, and the organ chip 2000 may be combined with a driving system 3000 into a whole through a connection base 5000, and the driving system 3000 is connected to a display control device 3100, and the control operation is performed through the display control device 3100.

The invention also provides a method suitable for biological tissue culture and real-time monitoring, which comprises the following steps of:

s110: using a biological 3D printer to construct a biological 3D printed tissue for testing/testing; wherein, the printing is directly carried out in the organ chip culturing room or carried out on the transferring unit and then placed in the organ chip culturing room;

s120: and culturing the test tissue according to the working mode selected by the user by receiving the working mode selected by the user.

Specifically, after receiving the working mode selected by the user, positive pressure air source and negative pressure air source are controlled to output positive pressure air flow and negative pressure air flow to the air channel distribution device, so that the air channel distribution device leads the positive pressure air flow and the negative pressure air flow to the air channel of the organ chip to enable the culture medium to move.

Specifically, after receiving a liquid feeding mode selected by a user, controlling a culture medium in a liquid storage tank or a culture chamber to enter a micro-channel;

after receiving the user-selected alternating pattern, controlling the flow of the culture medium in the organ-chip main body in a clockwise or counterclockwise alternating manner;

after receiving the circulation mode selected by the user, controlling the culture medium in the organ-chip main body to flow in a clockwise manner or a counterclockwise manner;

After receiving the step mode selected by the user, the circulation direction of the current culture medium flow is adjusted to be the previous flow direction or the next flow direction.

S130: and in the culture process, detecting the test tissue to obtain detection data.

Several examples of practical applications of the system and method for biological tissue culture and real-time monitoring provided by the present invention are described below.

Example 1

Step 1, taking a transfer unit 2100, printing biological 3D printing tissue with liver cells in the transfer unit by using a biological 3D printer 1000, and placing the transfer unit into a culture medium (DMEM culture medium+10% fetal bovine serum+1% green streptomycin) for 20 days, wherein the culture medium is replaced every two days;

step 2, a transfer unit 2100 is taken, a biological 3D printer 1000 is used for printing biological 3D printing tissue with tumor cells in the transfer unit, and the transfer unit is placed into a culture medium (DMEM culture medium+10% fetal calf serum+1% green streptomycin) for 20 days, and the culture medium is replaced every two days;

step 3, placing the liver cell-containing biological 3D printing tissue and the tumor cell-containing biological 3D printing tissue into a transfer unit 2100, and mounting the transfer unit 2100 to two different culture chambers on an organ chip 2000;

Step 4, placing the organ chip 2000 into the connection base 5000, closing the base cover 5010 to insert the air guide connector 5042 into the air pipe interface;

step 5, connecting a quick connector 3400 in the driving system 3000 by using a pipeline 3500, and connecting the driving system 3000 with a connecting base 5000;

step 6, turning on a switch 3120, waiting for the indication lamp 3110 to be turned on, using a touch screen 3130 to enter a driving system 3000 for selecting a liquid feeding mode, then adding a culture medium into a transfer unit 2100 by using a syringe or a pipette, allowing the culture medium to flow into a culture chamber, ending the liquid feeding mode after the whole micro-channel 2320 is filled with the culture medium, and returning to a working mode selection interface;

step 7, after the base cover 5010 is opened and the transparent top cover is placed on the organ chip 2000, the base cover 5010 is closed again to insert the air guide connector 5042 into the air pipe interface;

step 8, selecting a circulation mode in the driving system 3000, setting the circulation direction to be clockwise, setting the circulation frequency to be 0.5Hz, setting the positive pressure output value to be 150kPa, setting the negative pressure output to be 65kPa, and then starting the circulation mode;

step 9. The environment control system provides a constant temperature environment in this specific embodiment and controls the concentration of carbon dioxide, the connection base 5000 containing the organ chip 2000 is placed in an incubator, the connection is kept with the driving system 3000 outside the incubator through a pipe, the environment of the incubator is set to 37 ℃, and the concentration of carbon dioxide is 5%;

Step 10, analyzing biological information transmitted by the sensing chip through an analysis system to monitor the culture medium components in the micro-channel 2320 and deduce the growth condition of cells;

after 11.6 hours, opening a incubator door to take out a connection base 5000 containing an organ chip 2000, placing under an inverted optical microscope, observing the cell morphology on the biological 3D printing tissue in the incubator by using an observation window, opening a base cover 5010, removing a transparent top cover, sampling a culture medium in the incubator by using a pipette, and detecting cell proliferation of the sample;

step 12, adding the tested medicine into a culture room containing liver cell organism 3D printing tissue;

step 13, placing a transparent top cover on the organ chip 2000, then closing the base cover 5010 to insert the air guide joint 5042 into the air pipe interface and placing the connection base 5000 containing the organ chip 2000 into an incubator;

step 14, repeating the steps 10-13 for three times;

after 15.6 hours of step 10-11 is repeated, all medium in the culture chamber is removed by a pipette and new medium is added;

step 16, repeating the step 14 once after repeating the step 14-15 once;

after 17.6 hours, stopping the circulation mode in the driving system 3000, taking the connection base 5000 containing the organ chip 2000 out of the incubator, opening the base cover 5010, removing the transparent top cover, taking out all the culture medium in the incubator as a sample, taking out the biological 3D printing tissue containing cells for further testing;

And 18. Summarizing the sensing record, the cell proliferation result, the cell growth state observation record and the biological slicing result of the biological 3D printing tissue obtained in the experimental process, and analyzing to obtain the growth states of liver cells and tumor cells on the biological 3D printing tissue in the experimental process, wherein if the addition of the drug inhibits the growth of the tumor cells and does not influence the growth of the liver cells, the drug is screened by the hepatotoxicity.

Example 2

Step 1, respectively placing 2 transfer units 2100 into 2 culture chambers on an organ chip 2000;

step 2, enabling a printing needle head to enter a culture room by adjusting a printing nozzle of the biological 3D printer 1000;

step 3, respectively printing the biological 3D printing tissue with liver cells and the biological 3D printing tissue with tumor cells on the transfer units 2100 in 2 culture chambers;

step 4. Adding a culture medium (DMEM medium+10% fetal bovine serum+1% green streptomycin) to the culture chamber, and performing the operations of steps 4 to 7 in example 1;

step 5. Culturing the cells for 20 days, during which medium replacement is performed every two days as in step 6 of example 1;

step 6. Performing steps 8-17 of example 1;

and 7. Summarizing the sensing record, the cell proliferation result, the cell growth state observation record and the biological slicing result of the biological 3D printing tissue obtained in the experimental process, and analyzing to obtain the growth states of liver cells and tumor cells on the biological 3D printing tissue in the experimental process, wherein if the addition of the drug inhibits the growth of the tumor cells and does not influence the growth of the liver cells, the drug is screened by the hepatotoxicity.

Example 3

Step 1. Step 1-10 in example 1 is performed;

step 2, stopping a circulation mode in a driving system 3000 when a culture medium flows through due to insufficient concentration of a marker or insufficient reaction speed of a sensing chip, switching to an alternating mode, setting 3 clockwise circulation and then stopping for 5 seconds for 2 anticlockwise circulation, wherein the circulation frequency is 0.5Hz, the positive pressure output value is 150kPa, the negative pressure output value is 65kPa, and then starting the alternating mode;

step 3. Steps 9 to 17 in example 1 are performed, wherein "stop circulation mode in drive system 3000" in step 17 is changed to "stop alternate mode in drive system 3000";

and 4. Summarizing the sensing record, the cell proliferation result, the cell growth state observation record and the biological slicing result of the biological 3D printing tissue obtained in the experimental process, and analyzing to obtain the growth states of liver cells and tumor cells on the biological 3D printing tissue in the experimental process, wherein if the addition of the drug inhibits the growth of the tumor cells and does not influence the growth of the liver cells, the drug is screened by the hepatotoxicity.

Example 4

Step 1. Step 1-8 in example 1 is performed;

step 2, when bubbles are found in the micro flow channel 2320, stopping a circulation mode in the driving system 3000, selecting a stepping mode, adjusting the circulation direction to be the direction that the bubbles in the micro flow channel layer 2300 can enter the culture chamber without passing through the fence-shaped valve 2360, and manually controlling the stepping mode to slowly introduce the bubbles into the culture chamber and discharge the bubbles through the top of the liquid level of the culture medium;

Step 3, exiting a stepping mode in the driving system 3000, selecting a circulation mode, setting the circulation frequency to be 0.5Hz, outputting a positive pressure of 150kPa, outputting a negative pressure of 65kPa, and then starting the circulation mode;

step 4. Performing steps 9-17 of example 1;

and 5. Summarizing the sensing record, the cell proliferation result, the cell growth state observation record and the biological slicing result of the biological 3D printing tissue obtained in the experimental process, and analyzing to obtain the growth states of liver cells and tumor cells on the biological 3D printing tissue in the experimental process, wherein if the addition of the drug inhibits the growth of the tumor cells and does not influence the growth of the liver cells, the drug is screened by the hepatotoxicity.

Example 5

Step 1. Performing steps 1-8 of example 1

Step 2, when bubbles are found in the dividing groove 3250 or the liquid storage groove 3240, stopping the circulation mode in the driving system 3000 and selecting the manual mode;

step 3, manually controlling the positive pressure and the negative pressure output by the driving mode to control the volumes of the dividing groove 2350 and the liquid storage groove 2340, and squeezing bubbles into the micro-channels 2320 from the dividing groove 2350 or the liquid storage groove 2340;

step 4, stopping the circulation mode in the driving system 3000 and selecting the stepping mode, adjusting the circulation direction to be the direction that bubbles in the micro-channel layer 2300 can enter the culture chamber without passing through the fence-shaped valve 2360, and manually controlling the stepping mode to slowly introduce the bubbles into the culture chamber and discharge the bubbles from the top of the liquid level of the culture medium;

Step 5, exiting a stepping mode in the driving system 3000, selecting a circulation mode, setting the circulation frequency to be 0.5Hz, outputting a positive pressure of 150kPa, outputting a negative pressure of 65kPa, and then starting the circulation mode;

step 6. Performing steps 9-17 of example 1;

Example 6

Step 1. Performing steps 1-14 of example 1;

after 2.6 hours, opening a incubator door to take out a connection base 5000 containing an organ chip 2000, placing under an inverted optical microscope, observing the cell morphology on the biological 3D printing tissue in the incubator by using an observation window, opening a base cover 5010, removing a transparent top cover, sampling a culture medium in the incubator by using a pipette gun, adding a sample into a detection area, or after an ELISA strip is clamped in the detection area, adding the sample into the ELISA strip, and adding a marker;

Step 2, placing the organ chip 2000 into an enzyme labeling instrument for analysis and test;

step 3. Taking the organ chip 2000 out of the microplate reader and performing steps 15-17 in example 1;

Example 7

Step 1. Performing steps 1-3 of example 1

Step 2, opening a driving system 3000, pumping the culture medium in the liquid storage tank 6100 into a pipeline, connecting the pipeline with the liquid inlet 2220 and the liquid outlet 2230, allowing the culture medium to enter the micro-channel 2320 through the liquid inlet 2220, leaving the micro-channel 2320 through the liquid outlet 2230 after filling all the culture chambers with the culture medium, and flowing into the waste liquid tank 6200 through the pipeline;

step 3, adjusting the driving system 3000 to fix the flow rate at 20 microliters/second;

step 4, placing the organ chip 2000 which comprises two transferring units 2100 and is covered with a transparent top cover into an incubator, and setting a temperature of 37 ℃ and a carbon dioxide concentration of 5% in a position of a culture chamber in which the transferring unit 2100 containing tumor cells is positioned at a position more upstream than a culture chamber in which the transferring unit 2100 containing liver cells is positioned in a flow channel of a culture medium;

Step 5, analyzing the biological information transmitted by the sensing chip through an analysis system to monitor the culture medium components in the micro-channel 2320 and deduce the growth condition of cells;

after 6.6 hours, the organ-a-chip 2000 is taken out of the incubator, the waste liquid pool 6200 is disconnected from the pipeline, and the culture medium flowing out of the liquid outlet 2230 in the pipeline is further analyzed;

step 7, adding a culture medium containing the tested medicines into a liquid storage tank 6100;

step 8, repeating the steps 5-6;

step 9. After 72 hours from the start of the experiment, the organ-chip 2000 was taken out of the incubator after stopping the driving system 3000;

and step 10, summarizing the sensing record, the cell proliferation result, the cell growth state observation record and the biological slice result of the biological 3D printing tissue obtained in the experimental process, analyzing and obtaining the growth states of liver cells and tumor cells on the biological 3D printing tissue in the experimental process, wherein the addition of the medicine does not influence the growth of the liver and the tumor cells, and concluding that the medicine is ineffective or can not be directly effective on the tumor.

Example 8

Step 1. Performing steps 1-3 of example 1

step 4, placing the organ chip main body which comprises two transferring units 2100 and is covered with a transparent top cover into an incubator, and enabling a culture chamber in which the transferring unit 2100 containing tumor cells is positioned to be positioned at a position more downstream than a culture chamber in which the transferring unit 2100 containing liver cells is positioned in a flow channel of a culture medium, wherein the temperature is set to 37 ℃, and the carbon dioxide concentration is set to 5%;

step 8, repeating the steps 5-6;

and step 10, summarizing the sensing record, the cell proliferation result and the cell growth state observation record obtained in the experimental process and analyzing the biological slicing result of the biological 3D printing tissue to obtain the growth states of the liver cells and the tumor cells on the biological 3D printing tissue in the experimental process. The addition of the drug inhibits the growth of tumor cells without affecting the growth of liver cells, and the experimental result obtained in example 7 proves that the drug can produce drug effect on tumors after being metabolized by liver and has no hepatotoxicity.

Example 9

step 2, taking another transfer unit 2100, printing biological 3D printing tissue with kidney cells in the transfer unit by using a biological 3D printer 1000, and placing the transfer unit into a culture medium (DMEM culture medium+10% fetal bovine serum+1% green streptomycin) for 20 days, wherein the culture medium is replaced every two days;

step 3, respectively placing a transfer unit 2100 containing the liver cell organism 3D printing tissue and a transfer unit 2100 containing the kidney cell organism 3D printing tissue into 2 culture chambers on the organ chip main body;

step 4. Performing steps 4-9 of example 1;

step 5, analyzing biological information transmitted by the sensing chip and glucose concentration through an analysis system to monitor the culture medium components in the micro-channel 2320 and infer the growth condition of cells;

after 6.6 hours, opening a incubator door to take out a connection base 5000 containing an organ chip 2000, placing under an inverted optical microscope, observing the cell morphology on the biological 3D printing tissue in the incubator by using an observation hole, opening a base cover 5010, removing a transparent top cover, sampling a culture medium in the incubator by using a pipette, and detecting cell proliferation of the sample;

Step 7, removing all the culture mediums in the culture chambers by using a pipette, and adding a culture medium containing 15mmol/L glucose;

step 8, placing a transparent top cover on the organ chip 2000, then closing the base cover 5010 to insert the air guide joint 5042 into the air pipe interface and placing the connection base 5000 containing the organ chip 2000 into an incubator;

step 9, repeating the steps 5-8 until the pathological changes of the liver cells or the kidney cells are found by observing or sensing the data;

step 10, stopping a circulation mode in a driving system 3000, taking out a connection base 5000 containing an organ chip 2000 from an incubator, opening a base cover 5010, removing a transparent top cover, taking out all culture mediums in the incubator as samples, and taking out a biological 3D printing tissue containing cells for further testing;

and 11. Summarizing the sensing record, the cell proliferation result, the cell growth state observation record and the biological slice result of the biological 3D printing tissue obtained in the experimental process, analyzing and obtaining the growth states of liver cells and kidney cells on the biological 3D printing tissue in the experimental process, and summarizing the induction reasons of diabetes on liver and kidney lesions.

Example 10

Step 2, a transfer unit 2100 is taken, a biological 3D printer is used for printing biological 3D printing tissue with tumor cells in the transfer unit, and the transfer unit is placed into a culture medium (DMEM culture medium+10% fetal bovine serum+1% green streptomycin) for 20 days, and the culture medium is replaced every two days;

step 3, respectively placing a transfer unit 2100 containing the liver cell biological 3D printing tissue and a transfer unit 2100 containing the tumor cell biological 3D printing tissue into two different culture chambers on an organ chip, and sealing the organ chip by using a transparent top cover;

step 4, placing the organ chip on a connection base 5000, and integrating the connection base 5000 and a driving system 3000 in the same machine;

step 5, connecting the driving system 3000 with the touch screen 3130, setting the driving system 3000 through the touch screen 3130, selecting a circulation mode, setting the flow rate to be 30uL/min, and disconnecting the driving system 3000 from the touch screen 3130 after the setting is completed;

step 9. The environment control system in this specific embodiment is an incubator capable of providing a constant temperature environment, controlling carbon dioxide concentration and having a wire hole on the door, placing the driving system 3000 with the organ chip and the connection base 5000 in the incubator, wherein the environment of the incubator is set to 37 ℃ and the carbon dioxide concentration is 5%;

after 11.6 hours, opening a incubator door to take out the organ chip 2000, placing under an inverted optical microscope, observing the cell morphology on the biological 3D printing tissue in the incubator, removing the transparent top cover, sampling the culture medium in the incubator by using a pipetting gun, and detecting the cell proliferation of the sample;

step 12. The tested drug is added into a culture chamber 3110 containing hepatocyte growth 3D printed tissue,

step 13, placing the transparent top cover on the organ chip, then opening the incubator, connecting the organ chip on the connection base 5000, and closing the incubator;

step 14, repeating the steps 10-13 for three times;

step 16, repeating the step 14 once after repeating the step 14-15 once;

after 17.6 hours, taking the driving system 3000 connected with the organ chip and the connecting base 5000 out of the incubator, removing the transparent top cover, taking out all culture mediums in the incubator as samples, taking out the biological 3D printing tissue containing cells for further testing, connecting the touch screen 3130 with the driving system 3000, and stopping the circulation mode;

The invention combines organ chip technology with biological 3D printing technology to improve the culture mode of cell wall-attached growth of the existing organ chip. Specifically, cells of organs required to be cultured by an organ chip are mixed in biological 3D printing ink, and biological 3D printing tissue with the cells is printed out for culture. The method increases the survival time of the tissue, avoids the cell shedding to block the flow passage of the organ chip, and provides a culture method which is more similar to the in-vivo growth environment, so that the model constructed by the organ chip more simulates the human body.

Meanwhile, the growth state and the growth environment of the biological 3D printing tissue can be effectively evaluated through direct observation and on-line monitoring of the organ chip, so that more data for analyzing the growth process of the biological 3D printing tissue or emergency record can be timely executed when the growth of the biological 3D printing tissue is problematic, and the loss is reduced. In addition, the organ chip can be directly applied to detection instruments such as an enzyme-labeled instrument and the like, and is convenient to operate.

Third, the invention can also detect the growth state and metabolite content of the biological 3D printing tissue on line in real time, and simultaneously realize the automation of organ chip tissue culture, drug screening and pathological research operation, and reduce the influence of human factors.

Fourth, the transfer unit provides a transfer means between the 3D printer and the organ chip, and technical difficulties in printing directly in the organ chip are avoided. The connecting base can be used for rapidly plugging and unplugging the air pipe, and the complicated operation of plugging and unplugging the air pipe by the pneumatic organ chip is reduced.

The relative steps, numerical expressions and numerical values of the components and steps set forth in these embodiments do not limit the scope of the present invention unless it is specifically stated otherwise.

Any particular values in all examples shown and described herein are to be construed as merely illustrative and not a limitation, and thus other examples of exemplary embodiments may have different values.

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 addition, in the description of embodiments of the present invention, unless explicitly stated and limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. 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 description of the present invention, it should be noted that the directions or positional relationships indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. are based on the directions or positional relationships shown in the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.

Finally, it should be noted that: the above examples are only specific embodiments of the present invention, and are not intended to limit the scope of the present invention, but it should be understood by those skilled in the art that the present invention is not limited thereto, and that the present invention is described in detail with reference to the foregoing examples: any person skilled in the art may modify or easily conceive of the technical solution described in the foregoing embodiments, or perform equivalent substitution of some of the technical features, while remaining within the technical scope of the present disclosure; such modifications, changes or substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention, and are intended to be included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims

1. A system for biological tissue culture and real-time monitoring, comprising: the biological 3D printer, the organ chip, the connecting base, the driving system and the auxiliary system; the organ chip is connected with the driving system through the connecting base;

the auxiliary system is used for monitoring the state of biological 3D printing tissue;

the organ chip comprises a transparent top cover, a transfer unit and an organ chip main body; the transfer unit is detachably embedded into the organ chip main body, the transparent top cover covers the organ chip main body, and the transfer unit is used for containing the biological 3D printing tissue;

the organ-chip main body further includes: the micro-channel layer is arranged between the hard top layer and the transparent bottom layer, and the sensing chip is in contact with a culture medium of the organ chip main body;

wherein the transfer unit is matched with at least one culture chamber in the hard top layer and at least one culture chamber in the micro-channel layer;

the auxiliary system includes: an observation system, and/or an analysis system, and/or a control system, and/or an environmental control system;

the environment control system provides proper temperature, humidity and carbon dioxide concentration for biological 3D printing tissue growth;

the driving system comprises a display control device, a control device and an air path distribution device; the control device is respectively connected with the display control device and the air path distribution device;

2. The system of claim 1, wherein the organ-chip main body is rectangular or hexahedral in shape.

3. The system of claim 1, wherein the connection base is adapted to be used with the organ-chip, the connection base being used in combination with the drive system by way of a connection, or the connection base being used in combination with the drive system as a single unit.

4. The system of claim 1, wherein the system further comprises: the driving system is connected with the organ chip through the updating system and is also used for driving the updating system to update the culture medium in the organ chip.

5. A method suitable for biological tissue culture and real-time monitoring, characterized in that it is applied to the system according to any one of claims 1 to 4, wherein it comprises:

and in the culture process, detecting the test tissue to obtain detection data.

6. The method of claim 5, wherein said incubating said test tissue in said organ-chip according to a user-selected mode of operation by receiving a user-selected mode of operation comprises:

7. The method of claim 6, wherein after the step of incubating the test tissue in the organ-chip according to the user-selected mode of operation by receiving the user-selected mode of operation, the method further comprises: