Growth of embryonal carcinoma cells in serum-free medium

Two mouse embryonal carcinoma cell lines, PCC.4 aza-1 and F9, have been grown in serum-free F-12 medium supplemented with Pedersen fetuin, insulin, transferrin, and 2-mercaptoethanol. This medium supports long-term growth of both cell lines. When these cells are transferred from medium containing serum to this serum-free medium, growth continues without any detectable lag. PCC.4 aza-1 grown in this medium for over 20 generations retains the capacity to differentiate in vivo. This medium appears to be a general serum-free medium for the growth of embryonal carcinoma cells.