Specific steps for growing hamster kidney cells in cell culture flasks

When culturing various cells, cell culture flasks are a kind of cell culture consumables that are used more frequently. They are mostly used for the culture of adherent cells, and the specifications range from 25ml to 225ml. The hamster kidney cell culture medium will use this consumable, and the specific steps of its culture are as follows:

1. Preparatory work for cell culture:

1) Prepare MEM medium; high-quality fetal bovine serum, 10%; double antibody 1%.

2) Culture conditions: Gas phase: air, 95%; carbon dioxide, 5%. Temperature: 37 degrees Celsius, incubator humidity of 70%-80%.

3) Freezing solution: 90% serum, 10% DMSO, ready-to-use.

2. Cell processing:

1) Recovery of cryopreserved cells: Quickly shake and thaw the cryopreserved tube containing 1 mL of cell suspension in a 37°C water bath, add it to a centrifuge tube containing 4-6 mL of complete medium and mix well. Centrifuge at 1000RPM for 3-5min, discard the supernatant, and resuspend the cells in complete medium. The cell suspension was then added to a cell culture flask containing 6-8 ml of complete medium and cultured at 37°C overnight. Cell growth and cell density were observed under a microscope the next day.

2) Cell passage: If the cell density reaches 80%-90%, it can be subcultured.

3) Cryopreservation of cells: After receiving the cells, it is recommended to freeze a batch of cell seeds in the first 3 passages of culture for subsequent experiments.

The above are the specific steps for culturing hamster kidney cells in cell culture flasks. In addition, when cells are cryopreserved, collect the digested cells into a centrifuge tube according to the process of cell passage, and use a hemocytometer to count them to determine the cryopreservation density of cells. The recommended freezing density of general cells is 1×106~1×107 viable cells/ml.