4 mm punch biopsies were explanted to isolate primary keratinocytes, which were maintained in Green medium on a feeder layer of lethally irradiated 3T3 mouse fibroblasts as described previously (Barrandon and Green, 1987 (link)). For experiments, keratinocytes were grown in 0.06 mM CaCl2 EpiLife medium (Invitrogen) to 70–80% confluence and then incubated in 1.2 mM CaCl2 EpiLife medium for up to 48 hours to induce formation of desmosomes.
Duchatelet S., Boyden L.M., Ishida-Yamamoto A., Zhou J., Guibbal L., Hu R., Lim Y.H., Bole-Feysot C., Nitschké P., Santos-Simarro F., de Lucas R., Milstone L.M., Gildenstern V., Helfrich Y.R., Attardi L.D., Lifton R.P., Choate K.A, & Hovnanian A. (2018). Mutations in PERP cause dominant and recessive keratoderma. The Journal of investigative dermatology, 139(2), 380-390.
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