Preparation of mouse embryonic fibroblast (MEF) feeder plates for hPSC cultures v1
Year: 2022
Type: preprint
Abstract: This protocol describes the preparation of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell (hPSC) culture. Protocol overview 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 this protocol, 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 (start at step 4) or frozen stocks of irradiated or Mitomycin C inactivated MEFs can be used to prepare hPSC feeder cells. 3. The indicated MEF density are recommended starting densities and might have to be adjusted for each hPSC line and hPSC medium formulation (KSR, serum-free versus serum-containing medium). 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: W4210987414