Perfusion type organoid culture device

Abstract

The utility model discloses a perfusion type organoid culture device. The perfusion type organoid culture device comprises a fluid transmission module, a culture module and a storage module, the fluid transmission module comprises a peristaltic pump, a fluid conveying pipeline, a fluid recovery pipeline and a flow control valve; the culture module comprises a waste liquid collection container and an organoid culture connector; and the storage module comprises a liquid supply storage container and a waste liquid storage container. According to the device, pollution introduced from the outside in the step of liquid pumping or liquid changing in the traditional perfusion culture process can be avoided, cells can obtain sufficient oxygen in the liquid flowing process, the operation is simple and convenient, and automatic regulation and control and high-throughput culture can be realized.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the field of biomedical engineering, more particularly to a perfusion type organoid culture device. BACKGROUND

[0002] Organoids are three-dimensional structures and physiological functions of organs (or tissues) in vivo under normal (or disease) conditions in vitro, which can be simulated in vitro after 3D culture. Organoids can well simulate the in vivo environment, and organoids can well simulate the in vivo environment, which is suitable for molecular and cellular biology analysis, provides a better solution for tumor research, drug screening, regenerative medicine and other fields between animal and cell levels, and has been widely used in functional tissue induction, disease model establishment, drug screening, anti-inflammatory test, clinical end research and other aspects of research, and has great application in basic research and transformation application.

[0003] The culture device of organoids is a research hotspot in recent years. The early perfusion type culture device of organoids is relatively simple, mainly some improvements are made on the basis of traditional cell culture device, a simple perfusion system is introduced, and basic medium flow can be realized, which can provide more stable nutrition supply and metabolic waste discharge environment for organoids, but there are certain limitations in fluid control precision and culture environment simulation.

[0004] In the traditional culture method, the nutrition and oxygen are usually provided by simple medium soaking or regular medium replacement. For organoids with certain volume and structure, this method is difficult to ensure that the internal cells obtain sufficient nutrition and oxygen, and is easy to cause the cells in the center of organoids to die due to lack of nutrition and accumulation of metabolic waste, which affects the growth and development of organoids, so that it cannot completely simulate the real physiological state in vivo. Some traditional culture methods need frequent manual operation, such as medium replacement and subculture, which not only is tedious, but also increases the risk of cell contamination. At the same time, it is difficult to ensure completely consistent experimental conditions during operation, and human factors have a great influence on experimental results. Moreover, the traditional culture method is usually small-scale and single-sample culture, which is difficult to realize high-throughput organoid culture and screening. In the experiment of drug screening, toxicity test and other experiments requiring a large number of organoid samples, the efficiency is low, which cannot meet the rapid and large-scale experimental demand.

[0005] For fluid control, in recent years, injection pumps or hydraulic pumps are often used, but these fluid delivery devices are complex and require high-precision operation and maintenance. Therefore, there is an urgent need for an organoid culture device which is easy to operate, can stably provide the required nutrition and oxygen, can avoid contamination and can be cultured in high throughput. UTILITY MODEL CONTENT

[0006] The utility model discloses a purpose at overcoming prior art's defects and insufficient, provide a kind of perfusion type organoid culture device.

[0007] Based on the above purpose, the technical scheme adopted by the utility model is as follows:

[0008] A kind of perfusion type organoid culture device, including fluid transmission module, culture module and storage module;

[0009] The fluid transmission module includes peristaltic pump, fluid delivery pipeline, fluid recovery pipeline and flow control valve;

[0010] The culture module includes waste liquid collection container and organoid culture connector;

[0011] The storage module includes liquid supply storage container and waste liquid storage container;

[0012] Liquid supply storage container is connected with the upper end of organoid culture connector by fluid delivery pipeline, and flow control valve is arranged in fluid delivery pipeline;

[0013] Waste liquid collection container is arranged at the position of waste liquid dropped from the lower end of collection organoid culture connector;Waste liquid collection container is connected with waste liquid storage container by fluid recovery pipeline;

[0014] Fluid delivery pipeline and fluid recovery pipeline transport fluid by peristaltic pump.

[0015] Further, waste liquid collection container is provided with lid, and organoid culture connector is arranged in waste liquid collection container;The position of liquid inlet of fluid recovery pipeline is lower than organoid culture connector.

[0016] Further, the organoid culture connector includes pipeline connector, columnar liquid storage area and culture area;Pipeline connector is arranged at the top and / or side wall of columnar liquid storage area;The bottom of columnar liquid storage area is connected with culture area by neck structure;The organoid culture connector is hollow through structure.

[0017] Further, the culture area is semispherical.

[0018] Further, the bottom of semispherical culture area is columnarly extended.

[0019] Further, the number of liquid supply storage container is more than 1;When the number of liquid supply storage container is more than 1, fluid delivery pipeline is multiple-in-one pipeline, or multiple liquid supply storage containers are connected with multiple pipeline connectors of organoid culture connector by multiple fluid delivery pipelines.

[0020] Further, the peristaltic pump controls the plurality of pipes, the device is provided with a plurality of groups of liquid supply storage containers, fluid conveying pipes, flow control valves, organoid culture joints, waste liquid collection containers, fluid recovery pipes and waste liquid storage containers; each component is connected in the above manner.

[0021] Further, the perfusion type organoid culture device further comprises a data acquisition and control module.

[0022] The data acquisition and control module collects data of the flow sensor of the peristaltic pump, and controls the flow control valve and the peristaltic pump according to the collected data, so that the flow data is controlled within a set range.

[0023] Further, the perfusion type organoid culture device further comprises a temperature monitoring module,

[0024] The temperature monitoring module comprises a temperature sensor, a culture medium constant temperature heating device and a constant temperature culture device.

[0025] The outer wall of the culture area of the organoid culture joint is provided with a temperature sensor.

[0026] The fluid constant temperature heating device heats and keeps warm the liquid supply storage container.

[0027] The constant temperature culture device heats and keeps warm the organoid culture joint.

[0028] Further, the culture medium constant temperature heating device comprises at least one of an electric heating device and a water bath heating device.

[0029] Further, the constant temperature culture device comprises a constant temperature incubator.

[0030] Further, the data acquisition and control module further collects data of the temperature sensor, and adjusts the culture medium constant temperature heating device and the constant temperature culture device, so that the data of the temperature sensor is controlled within a set range.

[0031] The beneficial effects of the utility model are as follows:

[0032] 1. In the utility model, the peristaltic pump realizes the transportation of liquid outside the pipe, and in the whole process, the liquid is in a clean environment, so that the pollution introduced from the outside in the traditional perfusion type culture process can be avoided.

[0033] 2. In the utility model, the accurate control of flow rate can ensure that the organoid cells obtain sufficient nutrients, and when the culture medium drops, the wide culture area can make the cells obtain sufficient oxygen, and the problem that the traditional culture mode cannot obtain sufficient oxygen is avoided.

[0034] 3. The utility model discloses simple operation can be in the external control flow rate and temperature, avoid the pollution of complex operation introduction, and can realize automatic control through data acquisition and control module, avoid the error of human.

[0035] 4. The peristaltic pump can control multiple groups of pipelines, and high-throughput culture can be carried out through multiple groups of culture modules. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1 It is a schematic view of the perfusion type organoid culture device in example 1.

[0037] Figure 2 It is a schematic view of the organoid culture connector in example 1.

[0038] Figure 3 It is a schematic view of the organoid culture connector in example 1.

[0039] The serial number in the figure is explained: 1. Liquid supply storage container; 2. Fluid delivery pipeline; 3. Peristaltic pump; 4. Flow control valve; 5. Organoid culture connector; 6. Waste liquid collection container; 7. Fluid recovery pipeline; 8. Waste liquid storage container; 501. Pipeline connector; 502. Columnar liquid storage area and 503. Culture area. DETAILED DESCRIPTION

[0040] The utility model makes further detailed description in combination with the drawings and examples, but the implementation mode of the utility model is not limited to this.

[0041] Example 1

[0042] Reference Figure 1 A perfusion type organoid culture device, comprising a fluid transmission module, a culture module and a storage module;

[0043] The fluid transmission module comprises a peristaltic pump (3), a fluid delivery pipeline (2), a fluid recovery pipeline (7) and a flow control valve (4);

[0044] The culture module comprises a waste liquid collection container (6) and an organoid culture connector (5);

[0045] The storage module comprises a liquid supply storage container (1) and a waste liquid storage container (8);

[0046] The liquid supply storage container (1) is connected to the upper end of the organoid culture connector (5) through the fluid delivery pipeline (2), and the flow control valve (4) is arranged in the fluid delivery pipeline (2);

[0047] The waste liquid collection container (6) is arranged at a position where the waste liquid drops from the lower end of the organoid culture connector (5); the waste liquid collection container (6) is connected to the waste liquid storage container (8) through the fluid recovery pipeline (7).

[0048] The fluid delivery pipeline (2) and the fluid recovery pipeline (7) transport the fluid through the peristaltic pump (3).

[0049] In the embodiment, the organoid culture connector (5) is as shown in Figure 2 The organoid culture connector (5) is a hollow through structure, and the liquid can flow from the pipeline connector (501) through the columnar liquid storage area (502) and out of the culture area (503).

[0050] In the embodiment, the waste liquid collection container (6) is provided with a cover, and the organoid culture connector (5) is arranged in the waste liquid collection container (6); the liquid inlet of the fluid recovery pipeline (7) is arranged at a position lower than the organoid culture connector (5). The fluid is transported from the liquid supply storage container (1) to the pipeline connector (501) of the organoid culture connector (5) through the fluid delivery pipeline (2) provided with the flow control valve (4), then flows through the columnar liquid storage area (502) and the culture area (503) and drops from the culture area (503) into the waste liquid collection container (6), and finally is transported to the waste liquid storage container (8) through the fluid recovery pipeline (7). The peristaltic pump is used to transport the liquid outside the pipeline, and in the whole process, the liquid is in a clean environment, so that the pollution from the outside during the steps of pumping or replacing the liquid in the traditional perfusion culture process can be avoided.

[0051] The device is used for perfusion culture of organoids, and includes the following steps:

[0052] The fluid delivery pipeline (2) connected to the organoid culture connector (5) is pulled out, the organoid cells are connected to the columnar liquid storage area (502) of the organoid culture connector (5), and the pipeline connector (501) of the fluid delivery pipeline (2) and the organoid culture connector (5) is reconnected.

[0053] The peristaltic pump is started, and the peristaltic pump (3) and the flow control valve (4) are adjusted to control the target flow rate. The liquid supply storage container (1) is filled with the culture medium required for the organoid cell culture. The culture medium flows through the fluid delivery pipeline (2) equipped with the flow control valve (4) under the action of the peristaltic pump (3), is transported to the pipeline joint (501) of the organoid culture joint (5), and flows into the columnar liquid storage area (502). The cells will move downward to the culture area (503) under the action of the fluid and gravity, and will be stably grown in the culture area under the adhesion force. As shown in Figure 3 , the culture medium will flow through the cells in the culture area (503) and drip into the waste liquid collection container (6). During the dripping of the culture medium, the wide culture area (503) can provide sufficient oxygen for the cells, avoiding the problem that sufficient oxygen cannot be obtained in the traditional culture method. The waste culture medium that drips into the waste liquid collection container (6) is transported to the waste liquid storage container (8) through the fluid recovery pipeline (7) under the action of the peristaltic pump, avoiding the immersion of the collected culture medium into the organoid cells.

[0054] In some embodiments, as shown in B of Figure 2 , the pipeline joint is arranged on the side wall of the columnar liquid storage area, and the liquid can also flow into the columnar liquid storage area through the pipeline joint on the side wall.

[0055] In some embodiments, as shown in A of Figure 2 , the culture area is semispherical; in some embodiments, as shown in C of Figure 2 , the bottom of the semispherical culture area can be extended in a columnar shape.

[0056] In some embodiments, the number of liquid supply storage containers (1) is more than one; when the number of liquid supply storage containers (1) is more than one, the fluid delivery pipeline (2) is a multi-in-one pipeline, or multiple liquid supply storage containers (1) are connected to multiple pipeline joints (501) of the organoid culture joint (5) through multiple fluid delivery pipelines (2).

[0057] In some embodiments, the peristaltic pump controls multiple pipelines, and the device is provided with multiple sets of liquid supply storage containers (1), fluid delivery pipelines (2), flow control valves (4), organoid culture joints (5), waste liquid collection containers (6), fluid recovery pipelines (7), and waste liquid storage containers (8); each component is connected in the above manner.

[0058] In some embodiments, the perfusion type organoid culture device further comprises a data acquisition and control module.

[0059] The data acquisition and control module collects data of the flow sensor of the peristaltic pump, and controls the adjustment of the flow control valve and the adjustment of the peristaltic pump according to the collected data, so that the flow data is controlled within a set range.

[0060] In some embodiments, the perfusion type organoid culture device further comprises a temperature monitoring module,

[0061] The temperature monitoring module comprises a temperature sensor, a culture medium constant temperature heating device and a constant temperature culture device.

[0062] The outer wall of the culture area (503) of the organoid culture connector (5) is provided with a temperature sensor.

[0063] The fluid constant temperature heating device heats and keeps the liquid supply storage container (1) at a constant temperature.

[0064] The constant temperature culture device heats and keeps the organoid culture connector (5) at a constant temperature.

[0065] In some embodiments, the culture medium constant temperature heating device comprises at least one of an electric heating device and a water bath heating device.

[0066] In some embodiments, the constant temperature culture device comprises a constant temperature incubator.

[0067] Further, the data acquisition and control module also collects data of the temperature sensor, and adjusts the culture medium constant temperature heating device and the constant temperature culture device so that the data of the temperature sensor is controlled within a set range.

[0068] The above specific embodiments are preferred embodiments of the utility model, and cannot limit the utility model, and any change or other equivalent replacement mode without departing from the technical scheme of the utility model is included in the protection scope of the utility model.

Claims

1. A perfusion organoid culture device, comprising: The device comprises a fluid transmission module, a culture module and a storage module. The fluid transmission module comprises a peristaltic pump, a fluid delivery pipeline, a fluid recovery pipeline and a flow control valve. The culture module comprises a waste liquid collection container and an organoid culture connector. The storage module comprises a liquid supply storage container and a waste liquid storage container. The liquid supply storage container is connected to the upper end of the organoid culture connector through the fluid delivery pipeline, and the fluid delivery pipeline is provided with a flow control valve. The waste liquid collection container is arranged at a position where the waste liquid drops from the lower end of the organoid culture connector, and the waste liquid collection container is connected to the waste liquid storage container through the fluid recovery pipeline. The fluid delivery pipeline and the fluid recovery pipeline transport fluid through the peristaltic pump.

2. The device according to claim 1, wherein The waste liquid collection container is provided with a cover, and the organoid culture connector is arranged in the waste liquid collection container; and the fluid inlet of the fluid recovery pipeline is arranged at a position lower than the organoid culture connector.

3. The device according to claim 1, wherein The organoid culture connector comprises a pipeline connector, a columnar liquid storage area and a culture area; the pipeline connector is arranged at the top and / or sidewall of the columnar liquid storage area; the bottom of the columnar liquid storage area is connected to the culture area through a neck-shaped structure; and the organoid culture connector is a hollow through structure.

4. The device according to claim 3, wherein The number of the liquid supply storage containers is more than one; when the number of the liquid supply storage containers is more than one, the fluid delivery pipeline is a multi-in-one pipeline, or a plurality of liquid supply storage containers are connected to a plurality of pipeline connectors of the organoid culture connector through a plurality of fluid delivery pipelines.

5. The device according to claim 1, wherein The peristaltic pump controls a plurality of pipelines, and the device is provided with a plurality of sets of liquid supply storage containers, fluid delivery pipelines, flow control valves, organoid culture connectors, waste liquid collection containers, fluid recovery pipelines and waste liquid storage containers; and each component is connected in the manner of claim 1.

6. The apparatus of claim 1, wherein, The device further comprises a data acquisition and control module. The data acquisition and control module collects data of a flow sensor of the peristaltic pump, and controls the flow control valve and the peristaltic pump according to the collected data, so that the flow data is controlled within a set range.

7. The apparatus of claim 1, wherein, The device further comprises a temperature monitoring module. The temperature monitoring module comprises a temperature sensor, a culture medium constant-temperature heating device and a constant-temperature culture device. The outer wall of the culture area of the organoid culture connector is provided with a temperature sensor. The fluid constant-temperature heating device heats and keeps warm the liquid supply storage container. The constant-temperature culture device heats and keeps warm the organoid culture connector.

8. The device according to claim 7, wherein The culture medium constant-temperature heating device comprises at least one of an electric heating device and a water bath heating device. The constant-temperature culture device comprises a constant-temperature incubator.

9. The apparatus of claim 1, wherein, The device further comprises a data acquisition and control module and a temperature monitoring module. The temperature monitoring module comprises a temperature sensor, a culture medium constant-temperature heating device and a constant-temperature culture device. The outer wall of the culture area of the organoid culture connector is provided with a temperature sensor. The fluid constant-temperature heating device heats and keeps warm the liquid supply storage container. The constant-temperature culture device heats and keeps warm the organoid culture connector. The data acquisition and control module collects data from the flow sensor of the peristaltic pump, and controls the flow control valve and the peristaltic pump according to the collected data, so that the flow data is controlled within the set range; The data acquisition and control module also collects data from the temperature sensor, and controls the medium constant temperature heating device and the constant temperature culture device, so that the data of the temperature sensor is controlled within the set range.

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