Generating iPSC Lines from Human Dermal Fibroblasts
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Corresponding Organization :
Other organizations : National Heart Lung and Blood Institute, National Institutes of Health, National Institute of Neurological Disorders and Stroke, National Human Genome Research Institute
Protocol cited in 12 other protocols
Variable analysis
- Transduction of human dermal fibroblasts (HDFs) with Human STEMCCA Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit
- Addition of E8 medium without transforming growth factor–β (TGF-β) supplemented with 1 μM hydrocortisone and 100 μM butyrate
- Formation of Tra-1-60–positive iPSC colonies
- Culturing HDFs in Dulbecco's modified Eagle's medium containing 10% fetal calf serum (FCS) and 1% penicillin-streptomycin
- Coating plates with Matrigel (BD Biosciences)
- Passaging iPSC colonies with 0.5 mM EDTA (K&D Medical)
- Maintaining iPSC lines in full E8 medium (STEMCELL Technologies)
- Control human dermal fibroblasts (HDFs) generated from explants of 4-mm punch biopsy skin specimen
- ACDC-derived human dermal fibroblasts (HDFs) generated from explants of 4-mm punch biopsy skin specimen
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