Cell culture, cryopreservation and resuscitation in CAR-T cell therapy, and contamination control during production

According to the latest global cancer burden data released by the World Health Organization's International Agency for Research on Cancer (IACR) in 2020, there were 4.

What is cell culture?

Cell culture refers to the extraction of cells from animals or plants and then culture in an artificial environment for scientific research

Basic equipment and reagents required for cells

To conduct studies that require cell culture work and to perform basic cell culture protocols, Table 1 and Table 2 below summarize some of the key equipment and some basic reagents

Basic reagents required for cell culture

Other equipment includes a suction pump in a laminar flow hood that makes it easy to remove medium and reagents from the cell culture vessel; Autoclaves for disinfecting equipment and reusable glassware; Syringes, needles and forceps; Timers, 75% ethanol spray bottles and tissue rolls for disinfection of surfaces and equipment; Tape and permanent marker pens for marking; Pipe racks and waste bins, etc

Basic reagents required for cell culture

Cell culture basics

For cell survival and growth, it is critical that the culture environment replicates the physiological environment

Optimal cell culture conditions for most mammalian cells

Primary and immortal cells

Primary cell culture is cells that are isolated directly from intact or isolated tissue or organ fragments and grown in a petri dish

Some cell lines have no limits on their longevity and have unlimited proliferative capacity

Evolution and theoretical cell yield from primary cells to continuous cell lines

Cell culture cryopreservation and resuscitation

Cell lines are a valuable resource, so it is critical

The best way to freeze-preserve cultured cells is to keep them in liquid nitrogen in complete medium in the presence of cryoprotectants such as DMSO

The exact freezing conditions depend on the cell line

To restore cell lines from liquid nitrogen storage, transport frozen vials to the cell culture area

Recommendations for contamination control during the production of cell therapy products

In the production of cell therapy products (e.

Abandonment-oriented process solutions

First, the optional use of a disposable process solution reduces the risk of microbial contamination and cross-contamination and meets the requirements of

Pollution prevention and control of production and cultivation equipment

In addition, for production and cultivation equipment, it is more necessary to strictly control

The CO2 incubator placed in the production environment of the product can provide stable incubation environment conditions for the cells and ensure the quality of the
product.
The opening and closing door and exhaust air of the incubator will have a certain impact on the surrounding clean environment where it is placed, it is recommended to pay attention to the selection of the front-end equipment, and pay attention to maintenance and control after the equipment enters the use environment, so as to ensure that the equipment will not cause damage
to the external clean environment.
Special attention is paid to the environmental recommendations inside the incubator equivalent to the B-class clean environment, so that the internal and external cleanliness level is consistent when opening and closing the door, and the exhaust particles emitted during the operation of the equipment should be controlled in the hardware design to meet the needs of
the use environment.