US 12,297,415 B2
Fully enclosed cell culture system
Xuejun Yu, Shanghai (CN); Peng Xu, Shanghai (CN); Yongjun Fang, Shanghai (CN); and Dongge Feng, Shanghai (CN)
Assigned to Huadao (Shanghai) Biopharma Co., Ltd., Shanghai (CN)
Filed by HUADAO (SHANGHAI) BIOPHARMA CO., LTD., Shanghai (CN)
Filed on Mar. 30, 2021, as Appl. No. 17/216,699.
Application 17/216,699 is a continuation in part of application No. PCT/CN2019/098474, filed on Jul. 31, 2019.
Claims priority of application No. 201910184785.5 (CN), filed on Mar. 12, 2019; and application No. 201920312979.4 (CN), filed on Mar. 12, 2019.
Prior Publication US 2022/0348851 A1, Nov. 3, 2022
Int. Cl. C12M 1/00 (2006.01); C12M 1/02 (2006.01); C12M 1/04 (2006.01); C12M 1/38 (2006.01); C12M 3/00 (2006.01)
CPC C12M 3/00 (2013.01) [C12M 1/02 (2013.01); C12M 1/04 (2013.01); C12M 1/38 (2013.01); C12M 41/14 (2013.01)] 7 Claims
OG exemplary drawing
 
1. A fully enclosed cell culture system, comprising at least the following:
an incubator (1);
an bioreactor (2) to perform cell culture, disposed in the incubator (1), wherein the bioreactor (2) includes a stirrer (2.5), and the stirrer (2.5) is disposed in the bioreactor;
a gas flow assembly (3), connected with the incubator (1), wherein the gas flow assembly (3) adjusts concentrations of oxygen and carbon dioxide in the incubator (1);
a liquid flow assembly (4), connected with the bioreactor (2), wherein the liquid flow assembly (4) adjusts a liquid flow in the bioreactor (2), filters metabolites produced during cell culture, and recovers finished cells;
a temperature adjustment assembly (5), wherein the temperature adjustment assembly (5) adjusts temperature in the incubator (1);
a central controller (6), connected to and controlling the gas flow assembly (3), the liquid flow assembly (4), and the temperature adjustment assembly (5);
wherein the liquid flow assembly (4) comprises:
a liquid inlet passage (4.1), connected with the bioreactor (2); wherein the liquid inlet passage (4.1) feeds liquid into the bioreactor (2);
a circulation passage (4.2), connected with the bioreactor (2); wherein the circulation passage (4.2) has a filter 4.2.3 with a membrane having a pore size of 0.2-1 micrometers, which enables waste water to pass while preventing cells from passing through, therefore, cells in the waste water re-enter the bioreactor (2);
a waste liquid passage (4.3), wherein the waste liquid passage (4.3) is connected with the circulation passage (4.2) to discharge the metabolites; and
a harvest passage (4.4), wherein the harvest passage (4.4) is connected with the bioreactor (2) to recover finished cells;
wherein the gas flow assembly (3) comprises:
an air passage (3.1), a carbon dioxide passage (3.2) and an oxygen passage (3.3), which are separated and respectively connected with the incubator (1) to deliver gas into the incubator (1) to form a gas mixture;
a mixed gas suction passage (3.4), connected with the incubator (1) and the bioreactor (2), wherein the mixed gas suction passage (3.4) delivers the gas mixture in the incubator (1) into the bioreactor (2);
an exhaust gas discharge passage (3.5), connected with the bioreactor (2), wherein the exhaust gas discharge passage (3.5) discharges exhaust gas generated during a cell culture process;
a gas concentration sensor segment (3.6), including an oxygen gas concentration sensor (3.6.1) and a carbon dioxide gas concentration sensor (3.6.2), wherein the oxygen gas concentration sensor (3.6.1) measures a real-time oxygen gas concentration value, and the carbon dioxide gas concentration sensor (3.6.2) measures a real-time carbon dioxide gas concentration value in the incubator (1), wherein the gas concentration sensor segment (3.6) provides detection information to the central controller (6);
wherein the central controller (6) comprises:
a gas concentration comparison unit,
wherein the gas concentration comparison unit compares a real-time oxygen gas concentration value CtO2 measured by the gas concentration sensor segment with a preset oxygen gas concentration value C0O2,
wherein the gas concentration comparison unit compares a real-time carbon dioxide gas concentration value CtCO2 measured by the gas concentration sensor segment (3.6) with a preset carbon dioxide gas concentration value C0CO2,
wherein CO2 is the difference between C0O2 and CtO2 and CCO2 is the difference between C0CO2 and CtCO2, as given by formulas (I) and (II):
CO2=C0O2−CtO2  (I)
CCO2=C0CO2−CtCO2  (II); and
a gas concentration switch control unit, controlling the opening and closing of the carbon dioxide passage (3.2), the oxygen passage (3.3), and the mixed gas suction passage (3.4),
wherein an on-off time of the oxygen passage (3.3) is adjusted according to the value of CO2, and an on-off time of the carbon dioxide passage (3.2) is adjusted according to the value of CCO2,
wherein the mixed gas suction passage (3.4) is opened, and gas in the incubator (1) is sucked into the bioreactor (2), when both CO2 and CCO2 fall within a preset range;
when the mixed gas suction passage (3.4) is closed when either CO2 or CCO2 does not fall within its corresponding preset range.