A multi-channel gas-liquid interface culture device
Patent Information
- Application Number
- CN202522504409.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-26
- Publication Date
- 2026-10-09
- Estimated Expiration
- 2035-11-26
AI Technical Summary
[0004]本实用新型的目的在于提供一种多通道气液界面培养装置，其解决了现有培养基气液不稳定以及循环流动差的问题
[0010]作为本实用新型的一种优选方案，还包括顶层，所述顶层设置在中间层上方，且顶层上开设有对应培养槽、循环泵回液口、循环泵吸入口的通孔，本方案设置顶层用于方便安装循环泵等部件。本实用新型的有益效果在于：本方案将多组培养槽底部连通，结合流体力学，引入循环泵循环吸出与泵入培养基实现液体循环，培养装置配备多组培养槽并行培养满足高通量实验需求，循环泵吸入口的深度呈梯度变化，以便于从不同高度抽吸培养液，实现液面高度的可控调节，解决了营养供给不均、代谢废物堆积的问题，进一步缩小类器官与体内组织的功能差距，支持长期稳定培养。
Smart Images
Figure CN224832739U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of cell culture, specifically to a multi-channel gas-liquid interface culture device. Background Technology
[0002] Gas-liquid interface culture is a key technology developed to address the limitations of traditional organoid culture and to simulate the physiological microenvironment in vivo. Early organoid culture mainly used immersion 3D culture, which could form three-dimensional structures, but it was difficult to reproduce the complex microenvironment in vivo. Most target tissues in vivo are naturally in a gas-liquid interface environment, such as respiratory epithelium, skin epidermis, alveoli, and intestinal mucosa. The cells of these tissues are in contact with gas on one side and liquid (such as tissue fluid and nutrients) on the other side. For organoids to achieve functional differentiation (such as mucus secretion in respiratory epithelium and keratinization in skin), this "gas-liquid two-phase contact" microenvironment must be replicated. A simple liquid or gas environment cannot meet the physiological needs of cells.
[0003] Currently available solutions employ a simple "liquid level adjustment method," which raises the liquid level by changing the volume of added culture medium and placing a porous support membrane (such as a Transwell chamber) to place the organoids above the liquid and below the gas, thus initially achieving gas-liquid contact. However, the liquid level is uncontrollable, and the evaporation of the culture medium during the culture process makes the gas-liquid ratio even more unstable. Without the nutrient supply and metabolic waste removal of a circulating liquid environment, the cells are difficult to survive and cannot form more complex organoid structures. Utility Model Content
[0004] The purpose of this invention is to provide a multi-channel gas-liquid interface culture device, which solves the problems of unstable gas-liquid mixture and poor circulation in existing culture media.
[0005] This utility model achieves the above objectives through the following technical solutions: A multi-channel gas-liquid interface culture device includes a culture layer, a porous membrane, and several groups of culture tanks formed in the culture layer. The bottoms of each group of culture tanks are connected. The porous membrane divides the culture tank into an upper chamber and a lower chamber. At least one upper chamber in each group of culture tanks is provided with several circulation pump inlets with varying depth gradients.
[0006] As a preferred embodiment of this utility model, the culture layer is divided into a bottom layer and an intermediate layer. The lower cavity is opened on the bottom layer, and the upper cavity is disposed through the intermediate layer. A connecting groove is opened between the lower cavities of each group of culture tanks. In order to realize multi-channel culture of multiple groups of culture tanks, this embodiment modularizes the culture layer and the porous membrane, and divides the culture layer into a bottom layer and an intermediate layer.
[0007] In a preferred embodiment of this utility model, the porous membrane is disposed between the bottom layer and the middle layer. In this embodiment, the porous membrane is disposed as a whole and sandwiched between the middle layer and the bottom layer, and different regions of the porous membrane separate the culture tank.
[0008] As a preferred embodiment of this utility model, the intermediate layer is also provided with a circulation pump return port corresponding to each group of culture tanks, for connecting each group of culture tanks so as to facilitate the circulation of the culture medium. The circulation pump return port and the circulation pump suction port are respectively located at both ends of each group of culture tanks.
[0009] As a preferred embodiment of this utility model, each group of culture tanks is equipped with a corresponding circulation pump, and each circulation pump inlet is equipped with a reversing valve and multiple hollow tubes. The hollow tubes are connected to circulation pump inlets of different depths. This embodiment controls the height of the culture medium by setting circulation pump inlets with gradually varying depths. Furthermore, this embodiment uses hollow tubes inserted into the circulation pump inlets to draw culture medium from the bottom of the circulation pump inlets. When the culture medium height decreases, the culture medium is no longer drawn.
[0010] As a preferred embodiment of this utility model, it also includes a top layer, which is positioned above the middle layer. The top layer has through holes corresponding to the culture tank, the return port of the circulating pump, and the suction port of the circulating pump. This top layer facilitates the installation of components such as the circulating pump. The beneficial effects of this utility model are: this design connects the bottoms of multiple culture tanks, and combined with fluid dynamics, introduces a circulating pump to circulate and pump in the culture medium to achieve liquid circulation. The culture device is equipped with multiple culture tanks for parallel culture to meet the needs of high-throughput experiments. The depth of the circulating pump suction port varies gradually to facilitate the suction of culture medium from different heights, achieving controllable adjustment of the liquid level. This solves the problems of uneven nutrient supply and accumulation of metabolic waste, further narrowing the functional gap between organoids and in vivo tissues, and supporting long-term stable culture. Attached Figure Description
[0011] Figure 1 This is a schematic diagram of the culture layer and porous membrane structure of this utility model; Figure 2 For the present utility model Figure 1 A sectional view; Figure 3 This is a perspective view of the middle layer of this utility model; Figure 4 This is a top view of the middle layer of this utility model; In the diagram: 1. Culture layer; 101. Intermediate layer; 102. Bottom layer; 2. Porous membrane; 3. Culture tank; 301. Upper tank cavity; 302. Lower tank cavity; 303. Connecting tank; 304. Circulation pump suction port; 4. Circulation pump return port; 5. Top layer. Detailed Implementation
[0012] The present application will now be described in further detail with reference to the accompanying drawings. It should be noted that the following specific embodiments are only used to further illustrate the present application and should not be construed as limiting the scope of protection of the present application. Those skilled in the art can make some non-essential improvements and adjustments to the present application based on the above application content.
[0013] Example 1
[0014] like Figure 1-4 As shown, a multi-channel gas-liquid interface culture device includes a culture layer 1, a porous membrane 2, and several groups of culture tanks 3 formed in the culture layer 1. The bottom of each group of culture tanks 3 is connected. The porous membrane 2 divides the culture tanks 3 into an upper chamber 301 and a lower chamber 302. At least one upper chamber 301 in each group of culture tanks 3 is provided with several circulation pump inlets 304 with varying depth gradients.
[0015] This scheme connects the bottoms of multiple culture tanks 3 and, combined with fluid mechanics, introduces a circulating pump to circulate the culture medium, achieving liquid circulation. The culture device is equipped with multiple culture tanks 3 for parallel culture to meet the needs of high-throughput experiments. The depth of the circulating pump inlet 304 varies in a gradient to facilitate the aspiration of culture medium from different heights, achieving controllable adjustment of the liquid level. This solves the problems of uneven nutrient supply and accumulation of metabolic waste, further narrowing the functional gap between organoids and in vivo tissues, and supporting long-term stable culture.
[0016] Specifically, by using circulation pump suction inlets 304 of different depths, the suction height of the circulation pump can be selected as needed. Furthermore, by setting a buffer device between the circulation pump and the circulation pump return port 4, the circulation pump draws the culture medium into the buffer device and then slowly returns it to the circulation pump return port 4. It should be noted that the return speed of the circulation pump return port 4 is lower than the suction speed of the circulation pump. Therefore, the height of the culture medium can be controlled by the height of the circulation pump suction inlet 304. When adjustment is needed, the circulation pump suction inlet 304 of other depths can be switched.
[0017] Preferably, the culture layer 1 is divided into a bottom layer 102 and an intermediate layer 101. The lower cavity 302 is formed on the bottom layer 102, and the upper cavity 301 is disposed through the intermediate layer 101. A connecting groove 303 is formed between the lower cavities 302 of each group of culture tanks 3. In order to realize multi-channel culture of multiple groups of culture tanks 3, this scheme modularizes the culture layer 1 and the porous membrane 2, dividing the culture layer 1 into a bottom layer 102 and an intermediate layer 101. The porous membrane 2 is disposed between the bottom layer 102 and the intermediate layer 101, and the porous membrane 2 is set as a whole, sandwiched between the intermediate layer 101 and the bottom layer 102. Different regions of the porous membrane 2 separate the culture tanks 3.
[0018] The intermediate layer 101 is also provided with a circulation pump return port 4 corresponding to each group of culture tanks 3, which is used to connect each group of culture tanks 3 so that the culture medium can form a circulation flow. The circulation pump return port 4 and the circulation pump suction port 304 are respectively located at both ends of each group of culture tanks 3.
[0019] Each culture tank 3 is equipped with a corresponding circulation pump, and each circulation pump inlet 304 is equipped with a reversing valve and multiple hollow tubes. The hollow tubes are connected to circulation pump inlets 304 at different depths. This scheme controls the height of the culture medium by setting circulation pump inlets 304 with gradually varying depths. Furthermore, this scheme inserts hollow tubes into circulation pump inlets 304 to draw culture medium from the bottom of circulation pump inlets 304. When the culture medium height decreases, the culture medium is no longer drawn.
[0020] The culture device also includes a top layer 5, which is located above the middle layer 101. The top layer 5 has through holes corresponding to the culture tank 3, the return port 4 of the circulation pump, and the suction port 304 of the circulation pump, for easy installation of components such as the circulation pump.
[0021] Specifically, in this embodiment, the bottom layer 102 is divided into four independent channels. Each channel is equipped with six interconnected circular lower cavities 302 with a diameter of 14 mm and a depth of 5 mm for storing culture medium. The lower cavities 302 are connected by a connecting groove 303 with a width of 1 mm and a depth of 2 mm. A porous membrane 2 is laid on the bottom layer 102 and the middle layer 101. The porous membrane 2 is covered with a PDMS microporous film with a pore size of 5 μm (thickness 100 μm, porosity 35%). After being treated with plasma (power 100W, time 60s) to enhance hydrophilicity, it is laid flat between the bottom layer 102 and the middle layer 101 to ensure that the porous membrane 2 is completely adhered to the bottom layer 102 and the middle layer 101 without wrinkles or leakage. The intermediate layer 101 is equipped with 24 circular upper cavities 301 with a diameter of 14 mm for gas-liquid induction culture of organoids. The bottom culture medium permeates into the upper cavities 301 of the intermediate layer 101 through a microporous membrane to form a stable liquid film, allowing the organoids to be in contact with liquid nutrients and the gas environment at the top at the same time. The liquid outlet of the intermediate layer 101 is designed with multiple height restrictions on the liquid level. After the culture medium is drawn by the circulation pump, it is introduced into the inlet to realize the control of the liquid level during the gas-liquid induction culture process.
[0022] The top layer 5 is used to fix the pipes of the circulation pump, and is equipped with 10 rigid hollow tubes. Each hollow tube extends to a different depth into the middle layer 101. By changing the tubes used in conjunction with the circulation pump to draw culture medium, the liquid level can be controlled. The depths of each tube into the middle layer are 0.5mm, 1mm, 1.5mm, 2mm, 2.5mm, 3mm, 3.5mm, 4mm, 4.5mm, and 5mm, respectively. The inlet of the circulation pump (model: LabSmith C300, flow range 0.1-5mLin) is switched with the hollow tubes of the middle layer 101 at different depths via a reversing valve. The culture medium drawn from the circular culture tank 3 passes through the top inlet, through the middle layer 101, the porous membrane 2, and down to the bottom layer 102 to complete the circulation of the culture medium. By changing the reversing valve to change the hollow tubes drawn in, precise control of the liquid level (0-5mm) can be achieved.
[0023] This device allows for the adjustment of culture medium flow rate via a variable frequency circulating pump, supporting biomimetic simulation of peristaltic flow in intestinal organoids or respiratory fluid fluctuations in alveolar organoids. It employs a modular liquid distribution system, allowing each culture tank 3 to independently set liquid level, flow rate, and culture medium composition. For example, in drug toxicity testing, four different drug concentrations can be applied simultaneously to the same organoid model. The middle layer comprises 24 independent culture tanks 3 with a diameter of 14 mm, forming a 4×6 matrix liquid distribution network with the four independent channels (each channel containing six interconnected lower chambers) of the bottom layer 102. This design allows for the simultaneous gas-liquid induction culture of up to 24 organoids in a single operation, meeting the demands of high-throughput experiments.
[0024] The embodiments described above are merely examples of several implementations of this utility model, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of this utility model patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this utility model, and these modifications and improvements all fall within the protection scope of this utility model.
Claims
1. A multi-channel gas-liquid interface culture device, characterized in that, It includes a culture layer (1), a porous membrane (2) and several groups of culture tanks (3) opened in the culture layer (1). The bottom of each group of culture tanks (3) is connected. The porous membrane (2) divides the culture tank (3) into an upper chamber (301) and a lower chamber (302). At least one upper chamber (301) in each group of culture tanks (3) is provided with several circulation pump inlets (304) with varying depth gradients.
2. The multi-channel gas-liquid interface culture device according to claim 1, characterized in that, The culture layer (1) is divided into a bottom layer (102) and an intermediate layer (101). The lower cavity (302) is opened on the bottom layer (102), and the upper cavity (301) is arranged through the intermediate layer (101). A connecting groove (303) is opened between the lower cavities (302) of each group of culture tanks (3).
3. The multi-channel gas-liquid interface culture device according to claim 2, characterized in that, The porous membrane (2) is disposed between the bottom layer (102) and the intermediate layer (101).
4. The multi-channel gas-liquid interface culture device according to claim 2, characterized in that, The intermediate layer (101) is also provided with a circulation pump return port (4) corresponding to each group of culture tanks (3).
5. A multi-channel gas-liquid interface culture device according to claim 4, characterized in that, Each culture tank (3) is equipped with a circulation pump, and each circulation pump inlet (304) is equipped with a reversing valve and multiple hollow tubes, which are connected to circulation pump inlets (304) of different depths.
6. The multi-channel gas-liquid interface culture device according to claim 1, characterized in that, It also includes a top layer (5), which is located above the middle layer (101), and the top layer (5) has through holes for the corresponding culture tank (3), the return port of the circulation pump (4), and the suction port of the circulation pump (304).