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- W4309481815 abstract "Human epidermal keratinocytes (HEKs) typically undergo growth arrest after 15 - 30 population doublings (PDs) when maintained in traditional feeder-free culture medium formulations. To circumvent this early senescence, we developed DermaCult™ Keratinocyte Expansion Medium (KEM), a serum- and bovine pituitary extract (BPE)-free medium that promotes long-term growth (> 30 PDs) of HEKs while maintaining their air liquid interface (ALI) differentiation potential. To measure expansion over the lifespan of the culture, HEKs derived from neonatal (HEKn) or adult (HEKa) skin were thawed and seeded onto uncoated tissue cultureware at a density of 1 - 2.5 x 103 cells/cm2, and passaged once the cultures reached approximately 50 - 70% confluence. HEKs were assessed by qPCR and flow cytometry to detect the expression of cell markers of the basal and suprabasal epidermal layers. Differentiation potential was also assessed through ALI culture. DermaCult™ KEM supported 50 - 70 HEKa PDs and > 70 HEKn PDs before cell senescence (n = 4). HEKs cultured in DermaCult™ KEM maintained a cobblestone morphology, expressed basal cell markers (TP63, KRT14, ITGA6), and were devoid of suprabasal marker KRT10 (n = 4). In ALI culture, HEKs generated a stratified epithelium with the appropriate cellular architecture and expression of basal and suprabasal markers comparable to their in vivo counterparts (n = 2). Finally, keratinocytes obtained from human pluripotent stem cell (hPSC)-derived hair-bearing skin organoids and expanded in DermaCult™ KEM resembled those generated from primary HEKs, expressed basal keratinocyte markers (TP63, KRT14, KRT5), and sustained > 50 PDs before senescence (n = 4). In summary, DermaCult™ KEM is an improved serum- and BPE-free medium that supports the long-term expansion of primary and hPSC-derived HEKs while maintaining their downstream differentiation potential." @default.
- W4309481815 created "2022-11-28" @default.
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- W4309481815 date "2022-12-01" @default.
- W4309481815 modified "2023-09-25" @default.
- W4309481815 title "566 DermaCult™ Keratinocyte Expansion Medium, a Serum- and BPE-free Medium Supporting Long-term Feeder-free Expansion of Primary Human Epidermal and Pluripotent Stem Cell-Derived Keratinocytes" @default.
- W4309481815 doi "https://doi.org/10.1016/j.jid.2022.09.582" @default.
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