This was done as previously described11 (link). Briefly, the skin from euthanized neonatal mice of indicated genotypes was incubated with 250 units/mL dispase (Roche, cat. #04942078001) in KSFM medium (Thermo Fisher, cat. #17005-042) for 16 h at 4 °C to separate the epidermis from the dermis. The epidermis was then treated with accutase (CELLnTEC, cat. #CnT-accutase-100) for 30 min at 37 °C to release the keratinocytes. The isolated cells were then cultured in KSFM medium supplemented with 20 nM CaCl2, 25 μg/mL BPE and 0.25 ng/mL EGF (Thermo Fisher, cat. #37000-015).
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