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文献和实验3 and 4) and resuspend in a volume of fresh medium at least equivalent to the volume of trypsin. For cells that grow in clumps centrifuge and resuspend in a small volume and gently pipette to break up clumps. Under sterile conditions remove 100-200uL of cell
. No. D8537) using a volume equivalent to half the volume of culture medium. Repeat this wash step if the cells are known to adhere strongly. Pipette trypsin/EDTA (Prod. No. T4049) onto the washed cell monolayer using 1ml per 25cm2 of surface area
Cryopreservation of Cell Lines
. Pipette 1ml aliquots of cells into cyroprotective ampules that have been labeled with the cell line name, passage number, cell concentration and date. Place ampules inside a passive freezer e.g. Nalgene Mr. Frosty (Prod. No. C1562
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