a. Remove the frozen vial from the LN2 storage and transfer to 37℃; at once. b. When the cells are almost thawed (only a little chunk of ice), move the vials to ice bath. c. Transfer the cells to a 15ml sterile centrifuge tube and add 13-15ml culture medium. Centrifuge at 1000rpm for 5 minutes d. Remove the medium and resuspend the cell pellet with 8-10ml complete culture medium. Transfer the cells to a culture flask (T-25 flask). It is highly suggested to transfer 1ml of the culture from the T-flask into a 24 well plate. Place both T-25 flask and 24-well plate to a CO2 incubator. (*Optional step: If the cells grow not very well after recovering, mouse macrophage feeder cells could be added into 24-well plate and the amount is about 1*105/well.) e. Monitor the culture daily and pass the cells as needed.
Articles in this section
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