Cryopreservation and banking of mammalian英文.pdfVIP

PROTOCOL Cryopreservation and banking of mammalian cell lines Glyn N Stacey1 John R Masters2 1Division of Cell Biology and Imaging, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. 2Department of Surgery, Prostate Cancer Research Centre, Royal Free and University College London Medical School, 67 Riding House Street, London W1W 7EJ, UK. Correspondence should be addressed to J.R.M. (j.masters@ucl.ac.uk). Published online 4 December 2008; doi:10.1038/nprot.2008.190 s This protocol describes the principles and methods used for the preparation of cryopreserved cell stocks. Following these procedures l o c will ensure the availability of reproducible cultures for use within a single laboratory at different times and for different o t collaborating laboratories. Although the basic principle is simple, each cell line has characteristics that must to be borne in mind o r p when establishing and testing such stocks. The key requirements are reliable methods for culture, cryopreservation, characterization e r u and quality control. t a n / m o c INTRODUCTION . e r The need for cryopreserved cell banks and cell banking is added as a cryoprotectant to the culture medium, followed by u t a principles cooling at a rate of approximately 1 1C min1. Understanding n . w Maintaining a constantly growing culture of cells over many which elements of the cryopreservation process are critical to the w w months is bad practice. Any in vitro culture, whether prokaryotic successful pr