A novel fully automated three-dimensional cell culture device and its use method

Abstract

The present invention relates to a novel fully automated three-dimensional cell culture device, comprising: a master control device and disposable consumables matched with the master control device; the master control device comprises: a shell, a control element, a storage table and a carbon dioxide interface; the shell is further provided with a processing element, a digestion element, a gel mixing element and a culture element from left to right; the disposable consumables are placed on the storage table, comprising a processing chamber, a digestion chamber, a gel mixing chamber and a culture chamber connected from left to right through a conduit; a one-way air pressure valve is further provided in the conduit; a directional nozzle is further provided at the connection between the conduit and the culture chamber. The present invention can improve cell culture efficiency while saving labor costs, realizing standardized fully automated three-dimensional cell culture; it also has the advantages of simplicity, safety and high efficiency; the present invention can also realize high-throughput three-dimensional cell culture by adding modules, providing a standardized and large-scale solution for industrial production.

Description

Technical Field

[0001] The present invention relates to the field of cell culture technology, and in particular to a novel fully automated three-dimensional cell culture device and a method for using the same. Background Art

[0002] Cell culture is an essential process for bioengineering and is the most core and fundamental technology in biotechnology. Traditional artificial cell culture methods suffer from low efficiency, high risk of contamination, high error rates, and difficulty in implementing standardized quality control. Differing operator experience, techniques, and habits often lead to inconsistent cell states, resulting in poor reproducibility, stability, and uniformity of the culture process, thus affecting cell quality. Currently, the most reliable approach is to use automated systems to replace manual culture, which has become an industry trend. As demand for cell applications experiences explosive growth, more stringent requirements are being placed on improving cell production efficiency, effectively reducing production costs, and standardizing production quality standards to meet the huge market capacity and diverse needs of precision medicine. Currently, some automated, standalone devices on the market can only complete a specific step or section of the cell culture process, requiring human intervention to connect the entire production process, preventing full automation. At the same time, compared with traditional two-dimensional cell culture, three-dimensional cell culture has higher technical difficulty and more complicated operation steps, and is more difficult to standardize, automate, and scale. As a result, the cost of three-dimensional culture remains high, which greatly limits the further development of three-dimensional cell technology.

[0003] Currently, some automated, standalone devices on the market can only complete a single step or section of the cell culture process, requiring human intervention to connect the entire production process, rather than fully automated. Furthermore, compared to traditional two-dimensional cell culture, three-dimensional cell culture is technically more challenging and requires more complex procedures, making it more difficult to standardize, automate, and scale. This results in high costs for three-dimensional culture, significantly limiting the further development of this technology.

[0004] Therefore, there is an urgent need for a fully automated three-dimensional cell culture device that can easily replace culture consumables to achieve full process automation of three-dimensional cell culture. Summary of the Invention

[0005] The purpose of the present invention is to address the deficiencies in the prior art and provide a novel fully automated three-dimensional cell culture device and a method for using the same.

[0006] To achieve the above object, the technical solution adopted by the present invention is:

[0007] The first aspect of the present invention is to provide a novel fully automated three-dimensional cell culture device, comprising: a master control device and disposable consumables matching the master control device;

[0008] The master control device includes: a housing, a control element located at the upper portion of the housing, a storage platform located at the lower portion of the housing, and a carbon dioxide interface located at the top of the housing; the housing further includes, from left to right, a processing element, a digestion element, a gel mixing element, and a culture element; the processing element, the digestion element, the gel mixing element, and the culture element are electrically connected to the control element respectively;

[0009] The disposable consumables are placed on the storage table, including a processing chamber, a digestion chamber, a mixing chamber and a culture chamber connected from left to right in sequence through a conduit; a one-way air pressure valve is also provided in the conduit; a directional nozzle is also provided at the connection between the conduit and the culture chamber; the processing chamber, the digestion chamber, the mixing chamber and the culture chamber are respectively detachably and sealedly connected to the processing element, the digestion element, the mixing chamber and the culture element; each chamber and each element are connected with an elastic soft plastic material, which can ensure the sealing of the connection and can also buffer the impact of possible vibrations of each element (such as the mixing chamber).

[0010] Preferably, the top of the processing element is fixed to the upper part of the shell, the middle part can be telescopically adjusted up and down, and the bottom part extends into the processing chamber.

[0011] Preferably, the digestion element includes, from left to right, a respective independent first negative pressure component and a first liquid release component; the tops of the first negative pressure component and the first liquid release component are fixed to the upper part of the shell, and the bottoms extend into the digestion chamber; the first liquid release component stores digestive fluid.

[0012] Preferably, the glue mixing element includes, from left to right, a second negative pressure component and a second liquid releasing component, each of which is independent; the tops of the second negative pressure component and the second liquid releasing component are fixed to the upper part of the shell, and the bottoms extend into the glue mixing chamber; the second liquid releasing component stores matrix glue, and an oscillating temperature control component is also provided on it.

[0013] Preferably, the culture component includes, from left to right, a third negative pressure component, a gas release component and a third liquid release component, each of which is independent; the tops of the third negative pressure component, the gas release component and the third liquid release component are all fixed to the upper part of the shell, and the bottoms extend into the culture chamber; the gas release component is also connected to the carbon dioxide interface; the third liquid release component stores culture medium, and an oscillating temperature control component is also provided on it.

[0014] Preferably, a waste liquid pipe is further passed through the storage table; the waste liquid pipe is detachably and sealedly connected to the glue mixing bin and the culture bin respectively.

[0015] Preferably, a collecting tube is further passed through the storage platform; the collecting tube is detachably and sealedly connected to the culture chamber.

[0016] Preferably, a control valve is further provided in the waste liquid pipe and the collection pipe; the control valve is electrically connected to the control element.

[0017] A second aspect of the present invention is to provide a method for using the fully automated three-dimensional cell culture device as described above, comprising the following steps:

[0018] S1. Connect the carbon dioxide interface to the carbon dioxide storage tank; place the biological tissue in the processing chamber; place the disposable consumables on the storage table and seal them with each component of the master control device;

[0019] When the fully automated three-dimensional cell culture device is in operation, fully automated three-dimensional cell culture can be achieved through the control element;

[0020] S2, breaking the biological tissue into tiny tissue agglomerates by the processing element;

[0021] S3, the first negative pressure component pumps the tiny tissue agglomerates into the digestion chamber; the first liquid release component releases digestive fluid to digest the tiny tissue agglomerates into a cell suspension;

[0022] S4, the second negative pressure component pumps the cell suspension into the matrix gel mixing chamber; the second liquid release component releases the matrix gel, and the oscillating temperature control component mixes the cell suspension and the matrix gel;

[0023] S5. When the mixed cell suspension is pumped into the culture chamber by the third negative pressure component, the directional nozzle sprays the mixed cell suspension onto the inner wall of the culture chamber. After solidification, the third liquid release component releases the culture medium, and the gas release component releases carbon dioxide, thereby providing an external environment required for three-dimensional cell growth.

[0024] S6. After the culture is completed, the three-dimensional cells are collected and obtained through the collection tube; the waste liquid in the mixing chamber and the culture chamber is recovered and processed through the waste liquid pipe.

[0025] The present invention adopts the above technical solution, which has the following technical effects compared with the prior art:

[0026] The present invention is a fully automated three-dimensional cell culture method based on fluid control. By utilizing pressure control to move cells in various links of cell culture, it can improve cell culture efficiency while saving labor costs, and realize standardized fully automated three-dimensional cell culture. By using disposable and replaceable processing and culture consumables, the device cleaning steps and maintenance costs can be simplified, while avoiding cross-infection between samples, and has the advantages of simplicity, safety, and efficiency. The present invention can realize high-throughput three-dimensional cell culture by adding modules to process, culture and uniformly manage multiple samples at the same time, providing a standardized and large-scale solution for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Schematic diagram of the structure of the fully automated three-dimensional cell culture device of the present invention;

[0028] Figure 2 Schematic diagram of the structure of disposable consumables in the fully automated three-dimensional cell culture device of the present invention;

[0029] Among them, the figure marks include: shell 1; control element 2; storage table 3; carbon dioxide interface 4; processing element 51; digestion element 52; mixing element 53; culture element 54; waste liquid pipe 6; collection tube 7; processing chamber 8; digestion chamber 9; mixing chamber 10; culture chamber 11; one-way air pressure valve 12; directional nozzle 13. DETAILED DESCRIPTION

[0030] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts shall fall within the scope of protection of the present invention.

[0031] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0032] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but they are not intended to limit the present invention.

[0033] Example 1

[0034] like Figure 1-2 As shown, this embodiment provides a novel fully automated three-dimensional cell culture device, including: a master control device and disposable consumables matching the master control device;

[0035] The master control device includes: a shell 1, a control element 2 provided on the upper part of the shell 1, a storage table 3 provided on the lower part of the shell 1, and a carbon dioxide interface 4 provided on the top of the shell 1; the shell 1 is further provided with a processing element 51, a digestion element 52, a gel mixing element 53, and a culture element 54 from left to right; the processing element 51, the digestion element 52, the gel mixing element 53, and the culture element 54 are electrically connected to the control element 2 respectively; a waste liquid pipe 6 and a collection pipe 7 are also passed through the storage table 3; the waste liquid pipe 6 is detachably and sealedly connected to the gel mixing chamber 10 and the culture chamber 11 respectively; the collection pipe 7 is detachably and sealedly connected to the culture chamber 11; control valves are also provided in the waste liquid pipe 6 and the collection pipe 7; the control valves are electrically connected to the control element 2;

[0036] The disposable consumables are placed on the storage table 3, including a processing chamber 8, a digestion chamber 9, a glue mixing chamber 10, and a culture chamber 11 connected in sequence from left to right via a conduit; a one-way air pressure valve 12 is also provided in the conduit; a directional nozzle 13 is also provided at the connection between the conduit and the culture chamber 11; the processing chamber 8, the digestion chamber 9, the glue mixing chamber 10, and the culture chamber 11 are respectively detachably and sealedly connected to the processing element 51, the digestion element 52, the glue mixing element 53, and the culture element 54;

[0037] The top of the processing element 51 is fixed to the upper part of the housing 1, the middle part can be adjusted up and down, and the bottom part extends into the processing chamber 8;

[0038] The digestion element 52 includes, from left to right, a first negative pressure component and a first liquid release component, each of which is independent. The tops of the first negative pressure component and the first liquid release component are fixed to the upper part of the shell 1, and the bottoms extend into the digestion chamber 9. The first liquid release component stores digestive fluid.

[0039] The glue mixing element 53 includes, from left to right, a second negative pressure component and a second liquid release component, each of which is independent. The tops of the second negative pressure component and the second liquid release component are fixed to the upper portion of the housing 1, and the bottoms extend into the glue mixing chamber 10. The second liquid release component stores matrix glue and is also provided with an oscillating temperature control component.

[0040] The culture component 54 includes, from left to right, a third negative pressure component, a gas release component and a third liquid release component, each of which is independent; the tops of the third negative pressure component, the gas release component and the third liquid release component are all fixed to the upper part of the shell 1, and the bottoms extend into the culture chamber 11; the gas release component is also connected to the carbon dioxide interface 4; the third liquid release component stores culture medium, and an oscillating temperature control component is also provided on it.

[0041] Example 2

[0042] This embodiment provides a method for using the fully automated three-dimensional cell culture device as described in Example 1, comprising the following steps:

[0043] S1. Connect the carbon dioxide interface 4 to the carbon dioxide storage tank; place the biological tissue into the processing chamber 8; place the disposable consumables on the storage table 3 and seal them with each component of the master control device;

[0044] When the fully automated three-dimensional cell culture device is in operation, the control element 2 can realize fully automated three-dimensional cell culture;

[0045] S2, the processing element 51 breaks the biological tissue into small tissue agglomerates;

[0046] S3, the first negative pressure component pumps the tiny tissue agglomerates into the digestion chamber 9; the first liquid release component releases digestive fluid to digest the tiny tissue agglomerates into a cell suspension;

[0047] S4, the second negative pressure component pumps the cell suspension into the matrix gel mixing chamber 10; the second liquid release component releases the matrix gel, and the oscillating temperature control component mixes the cell suspension and the matrix gel;

[0048] S5. When the mixed cell suspension is pumped into the culture chamber 11 by the third negative pressure component, the directional nozzle 13 sprays the mixed cell suspension onto the inner wall of the culture chamber 11. After solidification, the third liquid release component releases the culture medium, and the gas release component releases carbon dioxide, thereby providing an external environment required for three-dimensional cell growth.

[0049] S6. After the culture is completed, the three-dimensional cells are collected and obtained through the collection tube 7; the waste liquid in the mixing chamber 10 and the culture chamber 11 is recovered and processed through the waste liquid pipe 6.

[0050] The present invention is a fully automated three-dimensional cell culture method based on fluid control. By utilizing pressure control to move cells in various links of cell culture, it can improve cell culture efficiency while saving labor costs, and realize standardized fully automated three-dimensional cell culture. By using disposable and replaceable processing and culture consumables, the device cleaning steps and maintenance costs can be simplified, while avoiding cross-infection between samples, and has the advantages of simplicity, safety, and efficiency. The present invention can realize high-throughput three-dimensional cell culture by adding modules to process, culture and uniformly manage multiple samples at the same time, providing a standardized and large-scale solution for industrial production.

[0051] The above description is only a preferred embodiment of the present invention and does not limit the implementation mode and protection scope of the present invention. For those skilled in the art, it should be aware that all solutions obtained by equivalent substitutions and obvious changes made using the description and illustrations of the present invention should be included in the protection scope of the present invention.

Claims

1. A fully automated three-dimensional cell culture device, characterized in that: include: Master control equipment and disposable consumables matching the master control equipment; The master control device comprises: a shell (1), a control element (2) provided at the upper part of the shell (1), a storage table (3) provided at the lower part of the shell (1), and a carbon dioxide interface (4) provided at the top of the shell (1); a processing element (51), a digestion element (52), a gel mixing element (53), and a culture element (54) are provided in the shell (1) from left to right; the processing element (51), the digestion element (52), the gel mixing element (53), and the culture element (54) are electrically connected to the control element (2), respectively; The disposable consumables are placed on the storage table (3), and include a processing chamber (8), a digestion chamber (9), a glue mixing chamber (10), and a culture chamber (11) connected in sequence from left to right via a conduit; a one-way air pressure valve (12) is also provided in the conduit; a directional nozzle (13) is also provided at the connection between the conduit and the culture chamber (11); the processing chamber (8), the digestion chamber (9), the glue mixing chamber (10), and the culture chamber (11) are detachably and sealedly connected to the processing element (51), the digestion element (52), the glue mixing element (53), and the culture element (54), respectively; A waste liquid pipe (6) is also passed through the storage platform (3); the waste liquid pipe (6) is detachably and sealedly connected to the glue mixing chamber (10) and the culture chamber (11). A collecting tube (7) is also passed through the storage platform (3); the collecting tube (7) is detachably and sealedly connected to the culture chamber (11).

2. The fully automated three-dimensional cell culture device according to claim 1, characterized in that: The top of the processing element (51) is fixed to the upper part of the shell (1), the middle part can be adjusted upward and downward, and the bottom part extends into the processing chamber (8).

3. The fully automated three-dimensional cell culture device according to claim 1, characterized in that: The digestion element (52) includes, from left to right, a first negative pressure component and a first liquid release component, each of which is independent; the tops of the first negative pressure component and the first liquid release component are fixed to the upper part of the shell (1), and the bottoms extend into the digestion chamber (9); digestive fluid is stored in the first liquid release component.

4. The fully automated three-dimensional cell culture device according to claim 3, characterized in that: The glue mixing element (53) includes, from left to right, a second negative pressure component and a second liquid releasing component, each of which is independent; the tops of the second negative pressure component and the second liquid releasing component are fixed to the upper part of the shell (1), and the bottoms extend into the glue mixing chamber (10); the second liquid releasing component stores matrix glue, and an oscillating temperature control component is also provided on it.

5. The fully automated three-dimensional cell culture device according to claim 4, characterized in that: The culture element (54) includes, from left to right, a third negative pressure component, a gas release component, and a third liquid release component, each of which is independent of the other. The tops of the third negative pressure component, the gas release component, and the third liquid release component are all fixed to the upper part of the shell (1), and the bottoms extend into the culture chamber (11). The gas release component is also connected to the carbon dioxide interface (4). The third liquid release component stores culture medium, and an oscillating temperature control component is also provided on it.

6. The fully automated three-dimensional cell culture device according to claim 5, characterized in that: Control valves are also provided in the waste liquid pipe (6) and the collection pipe (7); the control valves are electrically connected to the control element (2).

7. A method for using the fully automated three-dimensional cell culture device according to any one of claims 5 or 6, characterized in that the steps include: S1. Connect the carbon dioxide interface (4) to a carbon dioxide storage tank; place the biological tissue into the processing chamber (8); Placing the disposable consumables on the storage table (3) and sealingly connecting them to the components of the master control equipment respectively; S2, the processing element (51) breaks the biological tissue into small tissue clumps; S3, the first negative pressure component pumps the tiny tissue mass into the digestion chamber (9); The first liquid-releasing component releases digestive fluid to digest the tiny tissue agglomerates into a cell suspension; S4, the second negative pressure component pumps the cell suspension into the mixing chamber (10); The second liquid-releasing component releases the matrix gel, and the oscillating temperature-control component mixes the cell suspension with the matrix gel; S5, when the mixed cell suspension is pumped into the culture chamber (11) by the third negative pressure component, the directional nozzle (13) sprays the mixed cell suspension onto the inner wall of the culture chamber (11) in a directional manner, and after solidification, the culture medium is released by the third liquid release component, and the carbon dioxide is released by the gas release component, thereby providing an external environment required for three-dimensional cell growth; S6. After the culture is completed, the three-dimensional cells are collected and obtained from the collection tube (7); the waste liquid in the mixing chamber (10) and the culture chamber (11) is recovered and processed from the waste liquid pipe (6).

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