Resuspend the cell pellet: Use the recommended complete growth medium to resuspend the cell pellet.

Avoid alkaline medium: During recovery, avoid making the medium too alkaline. Before adding the vial contents, place the culture vessel in the incubator for at least 15 minutes to allow the medium to reach a pH of 7.0 to 7.6.

Incubate: Incubate the culture at 37°C in a suitable incubator. If using the medium described on the product sheet, use a 5% CO2 in air atmosphere.

Rinse the cell layer: Use a 0.25% (w/v) Trypsin-0.53mM EDTA solution to rinse the cell layer and remove any serum that contains trypsin inhibitor.

Avoid clumping: Do not shake or hit the flask to agitate the cells while waiting for them to detach.

Subcultivation ratio: Use a subcultivation ratio of 1:2 to 1:10.

Medium renewal: Renew the medium every 2 to 3 days.

Cell Image：

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