From: Nikolakis, G., Seltmann, H., Hossini, A. M., Makrantonaki, E., Knolle, J., & Zouboulis, C. C. (2015). Ex vivo human skin and SZ95 sebocytes exhibit a homoeostatic interaction in a novel coculture contact model. Experimental dermatology, 24(7), 497-502.

SZ95 sebocytes (24) were cultured in Sebomed basal medium (Biochrom, Berlin, Germany) supplemented with 10% foetal bovine serum (FBS), 50 lg/ml gentamycin, 10 ng/ml human epidermal
growth factor (EGF) and 1 mM CaCl₂ at 37°C and 5% CO₂ until reaching subconfluence. Normal human fibroblasts were isolated and expanded from skin of a healthy 21-year-old female volunteer and cultured in the same medium. Before the start of coculture experiments, SZ95 sebocytes and fibroblasts were resuspended in serum-free medium (Sebomed basal medium supplemented with 0.1% bovine serum albumin, 50 ug/ml gentamycin, 10 ng/ml human EGF, 1.5 mM CaCl₂, 1.5 x 10^-7 M linoleic acid and 10
6 M retinol) supplemented with a chemically defined serum substitute (Panexin-NTA; Pan Biotech, Aidenbach, Germany).
250 000 SZ95 sebocytes and 200 000 fibroblasts were seeded in 24-well plates and left overnight to attach at 37°C and 5% CO2. The next day the, wells were washed twice with PBS and treated with 400 and 600 ul of serum-free medium for direct contact and humoral contact coculture experiments, respectively.