WO2020000533A1 - Système de culture de cellules entièrement automatique hautement adaptatif et procédé associé - Google Patents

Système de culture de cellules entièrement automatique hautement adaptatif et procédé associé Download PDF

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Publication number
WO2020000533A1
WO2020000533A1 PCT/CN2018/096110 CN2018096110W WO2020000533A1 WO 2020000533 A1 WO2020000533 A1 WO 2020000533A1 CN 2018096110 W CN2018096110 W CN 2018096110W WO 2020000533 A1 WO2020000533 A1 WO 2020000533A1
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WO
WIPO (PCT)
Prior art keywords
unit
cells
culture
cell culture
automatic cell
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2018/096110
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English (en)
Chinese (zh)
Inventor
胡祥
刘沐芸
梁晓
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Cellauto Biological Automation Co Ltd
Shenzhen Beike Biotech Co Ltd
Original Assignee
Cellauto Biological Automation Co Ltd
Shenzhen Beike Biotech Co Ltd
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Application filed by Cellauto Biological Automation Co Ltd, Shenzhen Beike Biotech Co Ltd filed Critical Cellauto Biological Automation Co Ltd
Publication of WO2020000533A1 publication Critical patent/WO2020000533A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability

Definitions

  • the present application relates to a cell preparation system, and more particularly, to a highly adaptive fully automatic cell culture system and method thereof.
  • cell culture is also called cell cloning technology, and the formal term in biology is cell culture technology. Regardless of the entire biological engineering technology or one of the biological cloning technologies, cell culture is an indispensable process.
  • Cell culture itself is a large-scale cloning of cells.
  • Cell culture technology can transform a single cell into a simple single cell or rarely differentiated multi-cells. This is an essential part of cloning technology, and cell culture itself is the cloning of cells. A large number of cells or their metabolites are obtained through cell culture. Because biological products are derived from cells, it can be said that cell culture technology is the most core and basic technology in biotechnology.
  • the existing manual semi-automatic equipment on the market is aimed at local process steps in the full production process, and can only be produced for a single patient in the same period of time, which brings about the realization of industrialized cell production scale and effective reduction of production costs.
  • the benefits are limited.
  • the barriers established by foreign manufacturers in consumables, etc., have brought extremely high use costs to end-user users.
  • existing semi-automated equipment is usually customized and developed based on specific production processes, and it is difficult to meet the adaptability and scalability requirements of different production processes for different cell treatment schemes. Late The quality and quality control of large-scale production, it is difficult to provide systematic production and quality control data with existing semi-automated equipment.
  • the existing semi-automatic equipment still requires a lot of manual operation, and still requires the construction of a high-level clean room environment.
  • a highly adaptive fully automatic cell culture system including an isolation unit, a culture unit, a centrifugal transfection unit, and a collection unit;
  • the isolation unit is configured to perform separation and cleaning processing on a batch of cells and inoculate
  • the culture unit is used for preparing cultured cells
  • the centrifugal transfection unit is configured to perform virus centrifugal transfection and cell harvest on cells prepared in culture;
  • the collection unit is configured to perform detection on the acquired cells, and perform classification and transfer processing.
  • the system further includes an operation unit for operating cells in an isolation unit, a culture unit, a centrifugal transfection unit, and a collection unit, and the operation unit includes a six-degree-of-freedom GMP-compliant robot, A clamp and a dust-proof linear guide, wherein the clamp is connected to the six-degree-of-freedom GMP-compliant robot, and the six-degree-of-freedom GMP-compliant robot is disposed on the dust-proof linear guide.
  • the culture unit includes a CO 2 incubator and a culture bag storage module.
  • centrifugal transfection unit includes a centrifuge.
  • the assembly unit includes a detection module, a discharge module, and a waste storage module;
  • the detection module is configured to collect and detect basic quality data on the obtained cells
  • the discharging module is used for discharging the finished product
  • the waste storage module is configured to store used consumables and medicaments.
  • the system further comprises an environmental control unit
  • the environment control unit is configured to control the isolation unit, the culture unit, the centrifugal transfection unit, and the collection unit to be in a compliant and sterile environment of GMP.
  • the present application also provides a highly adaptive fully automatic cell culture method, which method includes:
  • the obtained cells are detected and classified and transferred out.
  • OKT3 is added, the density is adjusted, and cell seeding is performed.
  • a further technical solution thereof is: a step of performing culture preparation on the inoculated cells, including the following specific steps
  • the steps of performing centrifugal virus transfection and cell harvesting on the cultured cells include the following specific steps:
  • the contents of the packaging bag are washed and transferred to a new culture bag.
  • a highly adaptive fully automatic cell culture system of the present application by processing and seeding a batch system, culturing the seeded cells, and culturing the cultured cells Perform centrifugal transfection and detection output, and use the automatic control operation unit to run the cells in the isolation unit, culture unit, centrifugal transfection unit, and collection unit to achieve fully automatic cell culture, improve culture efficiency, and meet the requirements for mass production Production process adjustment and upgrade is highly adaptable, and The basic data of culturing cells strictly control the pass rate of cells.
  • FIG. 1 is a structural block diagram of a highly adaptive fully automatic cell culture system provided by a specific embodiment of the present application
  • FIG. 2 is a structural block diagram of a collection unit provided by a specific embodiment of the present application.
  • FIG. 3 is a flowchart of a highly adaptive fully automatic cell culture method provided by a specific embodiment of the present application.
  • FIG. 5 is a flow chart of culture preparation of inoculated cells provided by a specific embodiment of the present application.
  • FIG. 6 is a flowchart of virus centrifugation and cell harvesting of cells prepared in culture according to a specific embodiment of the present application
  • FIG. 7 is a flowchart of detecting and sorting out the acquired cells according to a specific embodiment of the present application.
  • a highly adaptive fully-automated cell culture system provided in this embodiment can be used to culture various cells such as red blood cells to achieve fully-automated cell culture and improve culture. Efficiency, and meet the needs of mass production, the production process adjustment and upgrade is highly adaptable.
  • this embodiment provides a highly adaptive fully automatic cell culture system, which includes an isolation unit 1, a culture unit 2, a centrifugal transfection unit 3, and a collection unit 4.
  • the isolation unit 1 is configured to perform a separation and washing process on a batch of cells and inoculate.
  • a culture unit 2 for culturing and preparing the seeded cells.
  • a centrifugal transfection unit 3 is configured to perform virus centrifugal transfection and cell harvesting on cells prepared in culture.
  • the collection unit 4 is configured to perform detection and classification transfer processing on the obtained cells.
  • the above-mentioned system further includes an operation unit 5 for operating cells in the isolation unit 1, the culture unit 2, the centrifugal transfection unit 3, and the collection unit 4.
  • the operation unit 5 includes a six-degree-of-freedom GMP compliance machine. A person, a clamp, and a dust-proof linear guide, the clamp is connected to the six-degree-of-freedom GMP-compliant robot, and the six-degree-of-freedom GMP-compliant robot is disposed on the dust-proof linear guide.
  • the operation unit 5 performs the transfer of patient cell samples, preparation consumables, etc. between different working modules, and participates in process operations such as adding liquid.
  • the cells are automatically transported in each culture unit 2, the cells are fully cultured to improve the culture efficiency, and the robot operation unit 5 based on a high degree of freedom and GMP certified is integrated to simulate human hands for core process equipment operations, with human hands During the process development process, the operation is highly adaptable to rapid process changes, and at the same time, it is highly adaptable to production process adjustments and upgrades during mass production.
  • the above-mentioned culture unit 2 includes a CO 2 incubator and a culture bag storage module. When the cells are cultured, they need to be cultured in a (: 0 2 incubator).
  • centrifugal transfection unit 3 includes a centrifuge, and performs operations such as virus (centrifugation) transfection and cell harvesting.
  • the above-mentioned collection unit 4 includes a detection module 41, a discharge module 42, and a waste storage module 43.
  • the detection module 41 is configured to collect and detect basic quality data on the obtained cells.
  • the discharging module 42 is used for discharging the finished product.
  • a waste storage module 43 is configured to store used consumables and medicaments.
  • the detection module 41 includes an automatic inverted microscope, a flow cytometer, and an optical cytometer. Collection of basic quality data during the preparation process, such as detection of total cells, cell proliferation rate, cell viability, transfection rate, cell subpopulations, etc.
  • the above-mentioned system further includes an environmental control unit 6.
  • the environment control unit 6 is configured to control the isolation unit 1, the culture unit 2, the centrifugal transfection unit 3, and the collection unit 4 in a GMP-compliant and sterile environment.
  • the ring The environmental control unit 6 includes an independent laminar flow module and an H202 sterilization module for each cabin.
  • the system is integrated with environmental control. Each cabin is sealed independently to form a class A dynamic laminar flow space.
  • the equipment can be installed and operated in a class D environment.
  • the system can be flexibly installed in hospitals, clinics, biological service companies, and pharmaceutical enterprise buildings. Highly adaptable.
  • the system also integrates a sterilization and disinfection system to meet the sterilization and disinfection needs of different production processes.
  • the above-mentioned system further includes a liquid operation unit 7.
  • the liquid operation unit 7 includes a bag-and-bottle liquid operation module, a large-capacity reagent solution operation module, and a small-capacity high-precision The liquid operation module, the bottle opening / closing module, and the magnet operation module.
  • the liquid operation unit 7 is configured to perform liquid operations with different capacities and different accuracy requirements in each process step.
  • the system also includes a refrigerator set including three temperature levels of -80 ° C, -20 ° C, and 4 ° C to provide environments such as low-temperature storage.
  • the system further includes an electric control unit 8.
  • the electric control unit 8 includes a high-performance industrial computing server, a PLC control module, various types of sensor networks, and a power supply module, etc., and provides power for each unit.
  • the system further includes a storage unit 9 for temporarily storing materials used in the production process in the system.
  • the system also includes units such as data collection, quality management, and remote customer service, adapting to the traditional production center production model, while creating a new distributed cell preparation production model, providing direct services close to the end user, and providing different production and business models. Highly adaptable.
  • the system, consumables, and supporting production processes can be uniformly reviewed and approved by the regulatory department, which is convenient for users to expand their business in multiple regions and increase production capacity.
  • the system can provide systematic operation and quality inspection reports in the production process, which is convenient for the regulatory department to efficiently monitor the final product of cell therapy.
  • the above-mentioned highly adaptive full-automatic cell culture system by processing and seeding a batch system, culturing the seeded cells, performing centrifugal transfection and detecting output on the cultured cells, using an automatic control operation unit 5
  • the cells are run in the isolation unit 1, culture unit 2, centrifugal transfection unit 3, and collection unit 4 to achieve fully automatic cell culture, improve the culture efficiency, and meet the high-adaptability to the production process adjustment and upgrade in the mass production process.
  • the basic data of the cultured cells are detected, and the qualified rate of the cells is strictly controlled.
  • this embodiment also provides a highly adaptive fully automatic cell culture method.
  • the method includes:
  • the steps of separating and washing the cells in batches and inoculating the cells include the following specific steps:
  • step S2 the step of culturing and preparing the seeded cells, includes the following specific steps:
  • S21 Sampling the inoculated cells to calculate the number and survival rate, and take pictures to save;
  • step S24 The cells in the culture preparation are tested.
  • step S3 the steps of performing virus centrifugation and cell harvesting on the cultured cells, include the following specific steps:
  • step S4 the steps of detecting and sorting out the acquired cells, include the following specific steps:
  • the cells are automatically transported in each culture unit 2, the cells are fully cultured to improve the culture efficiency, and the robot operation unit 5 based on a high degree of freedom and certified by GMP is integrated to simulate human hands for core process equipment operations, with human hands During the process development process, the operation is highly adaptable to rapid process changes, and at the same time, it is highly adaptable to production process adjustments and upgrades during mass production.
  • each cabin is independently sealed to form a Class A dynamic laminar flow space
  • the equipment can be installed and operated in a Class D environment
  • the system can be flexibly installed in hospitals, clinics, biological service companies, and pharmaceutical enterprise plants. Highly adaptable.
  • the system also integrates a sterilization and disinfection system to meet the sterilization requirements of different production processes.
  • the system, consumables, and supporting production processes can be uniformly reviewed and approved by the regulatory department, which is convenient for users to expand their business in multiple regions and increase production capacity.
  • the system can provide systematic operation and quality inspection reports in the production process, which is convenient for the supervision department to efficiently supervise the final product of cell therapy.
  • the above-mentioned highly adaptive full-automatic cell culture method includes processing and inoculating a batch system, cultivating the inoculated cells, performing centrifugal transfection and detecting output on the cultured cells, and using an automatic control operation unit 5
  • the cells are run in the isolation unit 1, culture unit 2, centrifugal transfection unit 3, and collection unit 4 to achieve fully automatic cell culture, improve the culture efficiency, and meet the high-adaptability to the production process adjustment and upgrade in the mass production process.
  • the basic data of the cultured cells are detected, and the qualified rate of the cells is strictly controlled.

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  • Genetics & Genomics (AREA)
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  • Cell Biology (AREA)
  • Medicinal Chemistry (AREA)
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  • Apparatus Associated With Microorganisms And Enzymes (AREA)
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Abstract

La présente invention concerne un système de culture de cellules entièrement automatique hautement adaptatif et son procédé associé, ledit système comprenant une unité d'isolement (1), une unité de culture (2), une unité de transfection centrifuge (3) et une unité de collecte (4) ; l'unité d'isolement (1) est utilisée pour séparer et laver une masse de cellules et effectuer l'inoculation ; l'unité de culture (2) est utilisée pour cultiver les cellules inoculées ; l'unité de transfection centrifuge (3) est utilisée pour effectuer la transfection centrifuge de virus et la récolte de cellules sur les cellules cultivées ; l'unité de collecte (4) est utilisée pour détecter les cellules récoltées et effectuer la classification et le transfert dessus. En utilisant une unité de transport (5) à commande automatique pour transporter les cellules vers l'unité d'isolement (1), l'unité de culture (2), l'unité de transfection centrifuge (3) et l'unité de collecte (4), réalisant la culture de cellules entièrement automatique, la présente invention améliore l'efficacité de culture, présente une adaptabilité élevée à l'ajustement et au passage à l'échelle supérieure du procédé de production dans la production de masse, détecte les données de base des cellules cultivées, et régule strictement le taux de qualification des cellules.
PCT/CN2018/096110 2018-06-26 2018-07-18 Système de culture de cellules entièrement automatique hautement adaptatif et procédé associé Ceased WO2020000533A1 (fr)

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CN201810666116.7A CN110643509A (zh) 2018-06-26 2018-06-26 一种高适应性全自动细胞培养系统及其方法
CN201810666116.7 2018-06-26

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CN111476517A (zh) * 2020-03-13 2020-07-31 青岛海尔生物医疗股份有限公司 一种细胞培养管理系统
CN121737026A (zh) * 2024-09-26 2026-03-27 北京赛桥未来生物科技有限公司 细胞分选及培养方法、装置、计算机设备及存储介质

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