{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T05:10:27Z","timestamp":1790745027191,"version":"4.1.0"},"reference-count":26,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2020,8,3]],"date-time":"2020-08-03T00:00:00Z","timestamp":1596412800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["currentprotocols.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["CP Stem Cell Biology"],"published-print":{"date-parts":[[2020,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Skin or hair loss (alopecia) may occur due to a wide variety of causes ranging from trauma to pathological processes including acquired or congenital causes. It would be ideal to replace them with immunologically compatible cells to avoid potentially exacerbating the condition. Deriving the replacement cells from human\u2010induced pluripotent stem cells (hiPSCs) allows for sufficient scale up and using hiPSCs as the choice of human pluripotent stem cells (hPSC) will ensure immunocompatibility. Here we offer a protocol for differentiating hiPSCs into keratinocyte progenitor cells (KPC) and keratinocytes employing all\u2010trans retinoic acid (ATRA) and <jats:sc>L<\/jats:sc>\u2010ascorbic acid, (L\u2010AA), bone morphogenic protein\u20104 (BMP4), and epidermal growth factor (EGF). We observed that the hiPSC\u2010derived KPCs express the same panel of markers as primary hair follicle bulge stem cells (HFBSCs), including CD200, integrin \u03b1\u20106 (ITGA6), integrin \u03b2\u20101 (ITGB1), the transcription factor P63, keratin 15 (KRT15), and keratin 19 (KRT19). If permitted to differentiate further, the hiPSC\u2010derived KPC lose CD200 expression and rather come to express keratin 14 (KRT14) indicating emergence of more mature terminally\u2010differentiated keratinocytes. The HFBSCs are transplantable for hair follicle (HF) restoration, and the keratinocytes may be transplantable for therapy for large burns or ulcers. \u00a9 2020 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Reprogramming of normal human skin fibroblasts into normal hiPSCs using episomal DNA cocktail<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Differentiation of hiPSCs into KPCs and keratinocytes<\/jats:p><jats:p><jats:bold>Alternate Protocol 2<\/jats:bold>: EBS formation protocol using AggreWell\u2122 plates (Antonchuk, 2013)<\/jats:p><jats:p><jats:bold>Support Protocol 1<\/jats:bold>: Passage hiPSC\u2010KPC<\/jats:p><jats:p><jats:bold>Support Protocol 2<\/jats:bold>: Immunocytochemistry (ICC)<\/jats:p><jats:p><jats:bold>Support Protocol 3<\/jats:bold>: Immunofluorescence staining of cells for flow cytometry (FC)<\/jats:p>","DOI":"10.1002\/cpsc.119","type":"journal-article","created":{"date-parts":[[2020,8,3]],"date-time":"2020-08-03T17:48:35Z","timestamp":1596476915000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Deriving Keratinocyte Progenitor Cells and Keratinocytes from Human\u2010Induced Pluripotent Stem Cells"],"prefix":"10.1002","volume":"54","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1810-6340","authenticated-orcid":false,"given":"Michel R.","family":"Ibrahim","sequence":"first","affiliation":[{"name":"Department of Dermatology, STD's and Andrology, Faculty of Medicine Minia University  Al\u2010Minya Egypt"},{"name":"Sanford Burnham Prebys Medical Discovery Institute  La Jolla California"},{"name":"Sanford Consortium for Regenerative Medicine  La Jolla 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