Maintenance and inactivation of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell culture v1
Year: 2022
Type: preprint
Abstract: This collection describes the maintenance and inactivation of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell (hPSC) culture. Collection overview Thawing of mouse embryonic fibroblasts (MEFs) for hPSC cultures Expansion of mouse embryonic fibroblasts (MEFs) for hPSC cultures Freezing of mouse embryonic fibroblasts (MEFs) for hPSC cultures Harvesting and irradiation of mouse embryonic fibroblasts (MEFs) for hPSC cultures Mitomycin C inactivation of mouse embryonic fibroblasts (MEFs) for hPSC cultures Preparation of mouse embryonic fibroblast (MEF) feeder plates for hPSC cultures A. Starting with frozen irradiated or Mitomycin C inactivated MEFs (optional) B. Starting with fresh irradiated or Mitomycin C inactivated MEFs General notes 1. Throughout these protocols, the term hPSC is used to collectively refer to both hiPSCs and hESCs. All described procedures have been tested and work equally well for hiPSCs and hESCs. 2. Either fresh or frozen stocks of irradiated or Mitomycin C inactivated MEFs can be used to prepare hPSC feeder cells. 3. The indicated MEF densities are recommended starting densities and might have to be adjusted for each hPSC line and hPSC media formulation (KSR, serum-free versus serum-containing media). 4. MEFs were obtained as described in Manipulating the Mouse Embryo: A Laboratory Manual, Third Edition (ISBN: 0879695919)
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W4211043233