Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells
作者： Anna E. Michalska
DOI: 10.1002/9780470151808.SC01C03S3
关键词:
摘要: To realize their potentials, embryonic stem (ES) cells must be maintained in optimal culture conditions that preserve pluripotency and self-renewal capacity. Mouse fibroblasts (MEFs) are used to prepare a feeder cell layer supports the growth of ES quality feeders is crucial for maintenance undifferentiated prolonged culture. The protocols provided this unit describe aspects isolation expansion MEFs established cells. Preparation mitotically inactivated (treatment with mitomycin C or γ-irradiation) also described. In addition, method counting numbers simple detection mycoplasma contamination by situ DNA staining provided. Methodology described has been tested real laboratory environment provides detailed information regarding resource time requirements as well critical parameters troubleshooting. Curr. Protoc. Stem Cell Biol. 3:1C.3.1-1C.3.17. © 2007 John Wiley & Sons, Inc. Keywords: embryonic cells; culture; mouse (MEFs); isolation expansion; feeder layer; MEFs freezing thawing; mycoplasma detection; protocols methods
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