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

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.