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Name: Curtis Okemwa OkeigaReg No: Btmb/059J/2012Title: Assignment 1: Protocol For Cryopreservation Course: Industrial Microbiology and BiotechnologyUnit: Cell, Tissue and Organ CultureDate: 5/12/14GENERAL PROTOCOL FOR THE CRYOPRESERVATION OF MAMMALIAN CELLS Introduction and Overview Mammalian cells can be cryopreserved (frozen) and maintained for many years (typically ten years or more) by careful preparation and strict attention to the details of the following guidelines. The two approaches outlined under this protocol can be used to successfully cryopreserve primary, normal, or continuous (immortal) cell types from either anchorage dependent or suspension growth systems. (This includes B and T lymphocytes separated from blood, transformed or not.) The basic tenant to follow in cryopreserving and subsequently recovering cells is to freeze slowly and thaw rapidly. Two of the most commonly used approaches to accomplish this are outlined here. Cryopreservation Medium The medium used to cryopreserve cells should always be the same formulation as that used to propagate the cells with the addition of fetal bovine serum (FBS) (in the case of serum-free cult

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