Human organoid complete culture system

Abstract

The utility model discloses a human-derived organ complete culture system which comprises a bottom plate, a pore plate layer, a bottoming reagent layer and a working solution layer which are sequentially arranged from bottom to top, the bottoming reagent layer and the working solution layer are respectively provided with a plurality of reagent grooves, and the reagent grooves of the bottoming reagent layer and the reagent grooves of the working solution layer are yin-yang grooves. The pore plate layer is composed of a plurality of layers of pore plates, the pore plates are covered with cover plates, vent pipes are arranged on the upper end faces of the cover plates, air outlets are formed in the pore plates, and the vent pipes and the air outlets are located at the two diagonal positions of the pore plates respectively. The breather pipe of the lower-layer pore plate is connected with the gas outlet of the upper-layer pore plate, so that the flow direction of gas between the upper-layer pore plate and the lower-layer pore plate is Z-shaped; the bottom plate and the lowermost-layer pore plate, the uppermost-layer pore plate and the bottoming reagent layer, and the bottoming reagent layer and the working solution layer are connected through buckles. By means of the layered design, the oxygen inlet and outlet channels are arranged on the pore plate layer, and all the layers have a synergistic effect and jointly maintain the culture environment of the organoids.

Description

Technical Field

[0001] This utility model relates to culture systems, and more particularly to a complete culture system for human organoids. Background Technology

[0002] Human organoid culture, as a cutting-edge technology in the life sciences, plays an irreplaceable role in disease research, drug development, and regenerative medicine. Organoids are three-dimensional tissue analogs formed in vitro through stem cell or progenitor cell culture. They can highly mimic the structure and function of organs in vivo, providing ideal experimental models for in-depth exploration of human physiological and pathological mechanisms, personalized drug screening, and the development of novel treatments.

[0003] Culture medium is a crucial factor in the culture of human organoids. In recent years, significant progress has been made in the technology of complete culture media for human organoids. Some traditional media have been modified to include various growth factors, cytokines, and nutrients, meeting the growth requirements of organoids to a certain extent. However, the culture effects of existing media are unstable. Different laboratories often produce different results when culturing the same type of organoids using the same culture medium formulation. This is mainly due to the poor stability of some components in the culture medium; for example, certain growth factors are easily inactivated during storage and use. Furthermore, the lack of uniformity in the production processes and quality control standards of culture media leads to inconsistent quality between different batches. Taking intestinal organoid culture as an example, intestinal organoids cultured from different batches of culture media exhibit significant differences in morphology, cellular composition, and function, making it difficult to replicate experimental results and hindering the widespread application of organoid technology in scientific research and clinical fields.

[0004] Furthermore, existing culture media are costly. Many components of these media are expensive, such as some recombinant growth factors and special nutrients, which not only increases research costs but also limits their widespread clinical application. In addition, the complex composition and stringent production conditions also increase the production cost of the culture media, making it difficult to popularize human organoid culture technology.

[0005] Numerous difficulties exist in attempting to address these challenges. On one hand, our understanding of the molecular mechanisms underlying organoid growth and development remains insufficient, making it difficult to precisely determine the optimal combination of culture medium components. On the other hand, developing new culture medium components or optimizing existing ones requires extensive experimental validation and long-term research, resulting in high costs and lengthy cycles. Furthermore, establishing unified quality control standards for culture media also faces challenges in terms of technology and regulatory oversight. Utility Model Content

[0006] This invention aims to solve the key problems existing in the prior art of complete human organoid culture media, and proposes a complete human organoid culture system.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] A complete human organoid culture system includes, from bottom to top, a base plate, a well plate layer, a bottom reagent layer, and a working solution layer. Both the bottom reagent layer and the working solution layer have several reagent tanks, with the bottom reagent tanks and the working solution tanks acting as opposite and opposite tanks to each other. The well plate layer consists of several well plates, each surrounded by a transparent plate. A cover plate is placed on the transparent plate, and the upper end of the cover plate has a venting tube for introducing gases required for cell growth into the well plate. The well plate has an outlet, with the venting tube and outlet located at two opposite corners of the well plate. The venting tube of the lower well plate is connected to the outlet of the upper well plate, causing the gas flow between the upper and lower well plates to form a Z-shape. The base plate and the bottom well plate, the top well plate and the bottom reagent layer, and the bottom reagent layer and the working solution layer are all connected by snap-fit ​​connections.

[0009] Furthermore, the air inlet of the vent pipe on the uppermost perforated plate is an external gas inlet, and the air outlet of the lowermost perforated plate is provided with an air outlet pipe to discharge the gas inside the perforated plate layer.

[0010] Furthermore, a reagent tank is provided on the left or right side of the upper end face of the bottom reagent layer, and correspondingly, a reagent tank is provided on the right or left side of the lower end face of the working liquid layer.

[0011] Furthermore, each perforated plate has external connecting supports at its four outer corners, and the upper and lower perforated plates are connected by these external connecting supports.

[0012] Working fluid layer: This is the part that directly contacts the organoid and contains various nutrients, growth factors, hormones, etc., required for cell growth. It provides energy and signaling molecules for the metabolism and growth of the organoid to support its normal physiological activities and developmental processes. For example, amino acids in it are the raw materials for protein synthesis, glucose provides energy for cells, and growth factors regulate important biological processes such as cell proliferation, differentiation, and survival.

[0013] The base layer (six grooves) primarily serves a supporting and immobilizing function, containing extracellular matrix components or other supporting materials. These components can mimic the structure and function of the extracellular matrix in vivo, providing a scaffold for organoid attachment and growth, while also helping to maintain cell morphology and polarity. For example, common extracellular matrix components such as collagen and laminin can promote cell adhesion and influence cell migration and differentiation.

[0014] 96-well plate: A 96-well plate is a container used for culturing organoids. Its multi-well design allows for the simultaneous culture of multiple samples, facilitating parallel experiments and comparative analysis. Each well can be considered an independent culture unit, enabling consistent control of culture conditions and facilitating high-throughput experimental research and screening.

[0015] The base plate serves as the bottom support for the entire culture medium structure, providing stability and structural integrity. It also functions as a connector or anchor to external devices for monitoring and controlling the culture environment, such as temperature and gas concentration. The snap-fit ​​design at the upper left corner of each layer likely facilitates assembly and disassembly, ensuring structural stability and sealing to prevent culture medium leakage or external contamination. It also allows for individual operation and maintenance of each layer.

[0016] This invention, through its layered design, provides a relatively stable, controllable, and in vivo-like environment for organoid culture, which helps improve the success rate and quality of organoid culture and provides strong support for the application of organoids in basic research, drug screening, and disease model construction. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of the structure of this utility model;

[0018] Figure 2 This is a schematic diagram of a single-layer perforated plate.

[0019] Figure 3 This is a diagram showing the usage state of this utility model;

[0020] In the diagram: 1. Base plate; 2. Orifice plate layer; 21. Vent tube; 22. Orifice plate; 23. Vent outlet; 24. Vent tube; 25. External connecting support; 26. Cover plate; 3. Bottom reagent layer; 31. Reagent tank; 4. Working liquid layer; 41. Reagent tank; 5. Snap-fit; 6. Incubator; 7. Gas circulation device; 8. Tube. Detailed Implementation

[0021] The technical solution of this utility model will be further described in detail below with reference to the accompanying drawings.

[0022] A complete human organoid culture system, such as Figure 1 and Figure 2As shown, the system includes, from bottom to top, a base plate 1, an orifice plate layer 2, a bottom reagent layer 3, and a working fluid layer 4. The bottom reagent layer 3 has several reagent tanks 31, and the working fluid layer 4 has several reagent tanks 41. The reagent tanks 31 and 41 are male and female tanks. The orifice plate layer 2 consists of several layers of orifice plates 22. The orifice plates 22 are surrounded by transparent glass plates (not shown in the figure). A cover plate 26 is placed on the transparent glass plates. The upper surface of the cover plate 26 is provided with a feature for guiding the orifice plates 22. 2. A ventilation tube 21 for introducing gas required for cell growth is provided. An outlet 23 is provided on the well plate 22. The ventilation tube 21 and the outlet 23 are located at two opposite corners of the well plate 22. The ventilation tube 21 of the lower well plate and the outlet 23 of the upper well plate are connected so that the gas flow direction between the upper and lower well plates is Z-shaped. The bottom plate 1 and the bottom well plate, the top well plate and the bottom reagent layer 3, and the bottom reagent layer 3 and the working liquid layer 4 are all connected by snap-fit ​​5.

[0023] The air inlet of the vent pipe 21 on the uppermost perforated plate is an external gas inlet, and the air outlet of the lowermost perforated plate is provided with an air outlet pipe 24 to discharge the gas inside the perforated plate layer.

[0024] A reagent tank 31 is provided on the left or right side of the upper end face of the bottom reagent layer 3, and correspondingly, a reagent tank 41 is provided on the right or left side of the lower end face of the working liquid layer 4.

[0025] Each perforated plate 22 has an external connecting post 25 at its four outer corners, and the upper and lower perforated plates 22 are connected by the external connecting posts 25. Specifically, one of the external connecting posts between the upper and lower perforated plates 22 is made solid and the other is made hollow, so that the solid connecting post can be inserted into the other hollow connecting post.

[0026] In actual use, such as Figure 3 As shown, the bottom reagent layer 3 and working solution layer 4 of the culture system are removed, and the well plate layer 2 together with the bottom plate 1 is placed in the incubator 6. The vent pipe 21 on the uppermost well plate is connected to the outlet of the gas circulation device 8 through the hose 8, and the vent pipe 24 on the lowermost well plate is connected to the inlet of the gas circulation device 8 through the hose 8, so that the well-mixed suitable gas enters the well plate through the gas circulation device 8. The incubator 6 provides the appropriate temperature and humidity required for cell culture. The connection between the gas circulation device 8 and the well plate layer 2 not only provides the gas environment required for cell culture, but also keeps the gas environment stable.

[0027] The above description is only a preferred embodiment of the present utility model and is not intended to limit the present utility model. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present utility model should be included within the protection scope of the present utility model.

Claims

1. A complete human organoid culture system, comprising, from bottom to top, a substrate, a well plate layer, a substrate reagent layer, and a working solution layer, characterized in that, Both the bottom reagent layer and the working solution layer are provided with several reagent tanks, and the reagent tanks of the bottom reagent layer and the reagent tanks of the working solution layer are mutually opposite tanks. The well plate layer is composed of several well plates. The well plates are surrounded by transparent plates, and a cover plate is placed on the transparent plates. The upper end of the cover plate is provided with a venting tube for introducing the gas required for cell growth into the well plate. The well plate is provided with an outlet. The venting tube and the outlet are located at two opposite corners of the well plate. The venting tube of the lower well plate is connected to the outlet of the upper well plate, so that the gas flow between the upper and lower well plates is in a Z-shape. The bottom plate and the bottommost well plate, the topmost well plate and the bottom reagent layer, and the bottom reagent layer and the working solution layer are all connected by snap-fit.

2. The complete human organoid culture system according to claim 1, characterized in that, The air inlet of the vent pipe on the uppermost perforated plate is an external gas inlet, and the air outlet of the lowermost perforated plate is equipped with an air outlet pipe to discharge the gas inside the perforated plate layer.

3. The complete human organoid culture system according to claim 1, characterized in that, A reagent tank is provided on the left or right side of the upper end face of the bottom reagent layer, and correspondingly, a reagent tank is provided on the right or left side of the lower end face of the working liquid layer.

4. The complete human organoid culture system according to claim 1, characterized in that, Each perforated plate has external connecting supports at its four corners, and the upper and lower perforated plates are connected by these external connecting supports.

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