Basics of Cell Culture

Working with cryopreservedcells• Vial from liquid nitrogen is placed into 37 C waterbath, agitate vial continuously until medium is thawed• Centrifuge the vial for 10 mts at 1000 rpm at RT, wipetop of vial with 70% ethanol and discard thesupernatant• Resuspend the cell pellet in 1 ml of complete mediumwith 20% FBS and transfer to properly labeled cultureplate containing the appropriate amount of medium• Check the cultures after 24 hrs to ensure that theyare attached to the plate• Change medium as the colour changes, use 20%FBS until the cells are established