CN-122003298-A - Automatic modular cell culture device with perfusion-based culture solution conveying function

CN122003298ACN 122003298 ACN122003298 ACN 122003298ACN-122003298-A

Abstract

The disclosed cell culture device comprises a cell culture module, a culture solution supply module, a culture solution storage chamber and a pair of culture solution pipes, wherein the cell culture module comprises a cell culture part formed by installing an orifice plate forming at least one culture space on a cassette, the culture solution supply module comprises a pump unit and forms a culture solution supply and discharge flow for the cell culture part, the culture solution storage chamber contains at least one culture solution chamber, the pair of culture solution pipes are made of flexible materials and are used for connecting the culture solution supply module and the cell culture module which are separated in space, a pipeline for forming the supply and discharge flow of the culture solution is formed, the cell culture module is placed in a CO 2 culture box for creating a warm environment, the culture solution storage chamber creates a low-temperature environment lower than normal temperature, and a pair of culture solution pipes exposed between the culture solution supply module and the cell culture module are in a normal-temperature environment.

Inventors

LI DONGXIAN
FANG JUNRONG
LI CHENGGAO
LI CHENGZHEN
Jin Dongcheng

Assignees

赛洛伊德株式会社

Dates

Publication Date: 20260508
Application Date: 20250424
Priority Date: 20240424

Claims (15)

1. A cell culture apparatus, wherein, The cell culture apparatus includes: A cell culture module including a cell culture part formed by installing an orifice plate forming at least one culture space on a cassette; the culture solution supply module comprises a pump unit for forming supply and discharge flow of culture solution to the cell culture part, a culture solution storage chamber for accommodating at least one culture solution chamber, and A pair of culture fluid pipes of flexible material connecting the culture fluid supply module and the cell culture module which are spatially separated, forming a pipeline for supplying and discharging the culture fluid, The cell culture module is arranged in a CO 2 incubator for creating a warm environment, the culture fluid storage chamber creates a low-temperature environment lower than normal temperature, and a pair of culture fluid pipes exposed between the culture fluid supply module and the cell culture module are in the normal temperature environment.
2. The cell culture apparatus according to claim 1, wherein, The pair of culture liquid pipes are a supply pipe and a discharge pipe, The pump unit sequentially pressurizes and supplies a predetermined amount of culture solution and gas to the supply pipe, thereby supplying the culture solution to the culture space, The inner diameter of the supply pipe has a diameter capable of preventing the culture medium from flowing backward due to gravity.
3. The cell culture apparatus according to claim 2, wherein, The pump unit sucks the culture liquid from the culture space by applying negative pressure to the discharge pipe, and recovers the culture liquid to a recovery chamber accommodated in the culture liquid storage chamber.
4. The cell culture apparatus according to claim 1, wherein, The cell culture apparatus includes a plurality of cell culture modules connected in parallel for one culture fluid supply module, By controlling the opening and closing of the valve modules provided in each of the plurality of cell culture modules, the one-to-one correspondence of the culture fluid supply modules to the cell culture modules to be transported is achieved during the time when the culture fluid supply and/or recovery flow is generated.
5. A culture solution supply module capable of supplying a culture solution to at least one culture space spatially separated and recovering the culture solution from the culture space, wherein, The culture medium supply module includes a pump unit for supplying and discharging a culture medium to and from the cell culture section, and a culture medium storage chamber for accommodating a recovery chamber and at least one culture medium chamber, The culture solution storage chamber creates a low-temperature environment lower than normal temperature, The pump unit supplies the culture liquid stored in the culture liquid chamber to each of the culture spaces through a supply pipe, and discharges the culture liquid to the recovery chamber through a discharge pipe.
6. The culture fluid supply module according to claim 5, wherein, The pump unit comprises a plurality of syringe pumps, The plurality of syringe pumps includes a gas syringe, at least one or more culture fluid syringes, and a recovery syringe.
7. The culture fluid supply module according to claim 6, wherein, The gas injector and at least one or more culture solution injectors are connected in parallel with the supply pipe, Check valves are provided upstream and downstream of the junction with the supply pipe, respectively.
8. The culture fluid supply module according to claim 7, wherein, The gas injector is arranged most upstream of the supply pipe, Check valves provided at the gas injector and the culture solution injector allow only a single direction of flow toward the culture space.
9. The culture fluid supply module according to claim 8, wherein, The gas injector is provided with an air filter upstream of the upstream check valve.
10. The culture fluid supply module according to claim 8, wherein, A predetermined amount of culture solution is introduced into the supply pipe through the suction and discharge strokes of the culture solution injector, The gas introduced into the supply pipe through the suction and discharge stroke of the gas injector conveys the culture solution introduced into the supply pipe to the culture space.
11. The culture fluid supply module according to claim 10, wherein, The plurality of syringe pumps includes a gas syringe and a plurality of culture fluid syringes, The culture medium is transferred between any one of the culture medium injectors and the gas injector at a staggered timing, so that the plurality of culture media to be transferred are transferred to the culture space in a state of being isolated by gas.
12. The culture fluid supply module according to claim 6, wherein, The recovery syringe is connected to the most downstream of the discharge tube, Check valves are provided upstream and downstream of the junction of the recovery syringe and the discharge tube, respectively, so as to allow flow in only a single direction toward the recovery chamber.
13. The culture fluid supply module according to claim 6, wherein the pump unit includes a plurality of pump actuators for linearly moving plungers of the syringe pumps to perform suction and discharge strokes.
14. A culture fluid supply module capable of supplying a culture fluid to at least one culture space spatially separated, wherein, The culture solution supply module includes: A pump unit including a plurality of syringe pumps for forming a flow of the culture medium supplied to the cell culture section, and a culture medium storage chamber for accommodating at least one or more culture medium chambers, The plurality of syringe pumps includes a gas syringe connected to the most upstream of the supply pipe, and at least one or more culture liquid syringes connected in parallel to the downstream side of the gas syringe, Check valves are provided upstream and downstream of the junction of the gas injector and each of the culture liquid injectors with the supply pipe, respectively, so as to allow only a single-directional flow toward the culture space, A predetermined amount of the culture solution is introduced into the supply pipe through the suction and discharge strokes of the culture solution injector, and the gas introduced into the supply pipe through the suction and discharge strokes of the gas injector transfers the culture solution introduced into the supply pipe to the culture space.
15. The culture fluid supply module of claim 14, wherein, The plurality of syringe pumps includes a gas syringe and a plurality of culture fluid syringes, The culture medium is transferred between any one of the culture medium injectors and the gas injector at a staggered timing, so that the plurality of culture media to be transferred are transferred to the culture space in a state of being isolated by gas.

Description

Automatic modular cell culture device with perfusion-based culture solution conveying function Technical Field The invention relates to an automatic cell culture device, in particular to a modular cell culture device, wherein a cell culture module and a culture solution supply module are connected through a pipeline network, the culture solution is automatically supplied and discharged, the cell culture device can be miniaturized to the extent that the cell culture module is only placed in a commercial CO 2 incubator, a very small amount of culture solution can be accurately conveyed through perfusion-based culture solution conveying, and the cell culture device can be supplied without deterioration in the conveying process. The present application claims priority rights of korean patent application No. 10-2022-0042443, month 05 of 2024, and korean patent application No. 10-2022-0042443, month 05 of 2024, which are incorporated in the present specification as part of the present specification in their entirety. Background Cell culture is an indispensable technique for studying and applying functions, characteristics, diseases, treatments, and the like of cells in various fields. However, cell culture generally requires a lot of time and effort, and particularly, there are many risk variables such as contamination, variation, etc. during cell culture, and thus, a very precise culture environment is also required. In addition, the quality and repeatability of cell culture are highly dependent on the skill and operation technique of operators, so that there is also a problem that it is difficult to ensure the reliability and consistency of experimental results. Under such circumstances, there has been an increasing demand for an automated cell culture apparatus in recent years, and many enterprises have been devoted to the development of a technique for automating a cell culture process by taking into account human and physical resources. The present invention has been made in view of the above-described problems, and is characterized by realizing environmental automation and automatically replacing a culture medium required for cell growth in a culture space in which cells can be cultured. In addition, the present invention aims to provide additional technical elements that are not obvious to those skilled in the art, in addition to the technical ideas mentioned above. Disclosure of Invention Problems to be solved by the invention In order to perform a cell culture process with a long time span, the cell culture module needs to be disposed in a space in a warm environment (usually about 37 ℃) suitable for cell growth, and the culture fluid supply module needs to be disposed in a space in a low-temperature environment (usually 4-8 ℃) to prevent the stored culture fluid from deteriorating. In this way, when considering means for maintaining the two spaces at different temperatures from each other, the present invention has a main object of moving an accurate amount of culture medium between a cell culture module and a culture medium supply module without wasting and deteriorating by physically separating a warm space for cell culture from a low-temperature space for culture medium storage and realizing perfusion-based culture medium transport to which an elongated culture medium tube and a gas-liquid interface transport mechanism are applied, because of heat transfer between the two spaces, which may cause derivative problems such as energy waste and moisture condensation. In addition, the most critical problem in the cell culture process is the cross contamination caused by the inflow of the contamination source from the outside, so that the cell culture apparatus must block the risk of contamination from the source during use, and further, in order to improve productivity and process efficiency, all the surfaces that may contact the culture solution and cells should be designed to be detachable from the apparatus so that the cells that have been previously cleaned and sterilized can be used in the culture operation. The present invention has been made in view of the above problems, and another object is to eliminate the washing and sterilization steps during the cultivation. For reference, examples of the surface to which the culture medium and the cells may contact may include a wall surface of a culture space in which the cells are cultured, a wall surface of a storage space in which the culture medium is stored, a wall surface of a pipe in which the culture medium moves, a pump and a valve for generating a flow, and the like. Further, an object of the present invention is to provide an automated cell culture apparatus in which a cell culture module and a culture fluid supply module are connected via a pipeline network to automatically supply and discharge a culture fluid, and which can be miniaturized to such an extent that only the cell culture module is placed in a commercial CO 2 incubator. In addition,