Efficiency of neural differentiation of mouse P19 embryonal carcinoma cells is dependent on the seeding density.

Abstract

Serum-free culture conditions for retinoic acid-induced neural differentiation of mouse P19 embryonal carcinoma cells were determined for future ex vivo retroviral gene transfer and brain transplantation studies. Neural differentiation of P19 cells was dependent on the seeding densities, and both neurons and astroglia differentiated efficiently at high seeding densities (2 x 10(4) and 5 x 10(4) cells/cm2) but not at low seeding density (1 x 10(4) cells/cm2). In addition, P19 cells cultured at 5 x 10(4) cells/cm2 showed neural differentiated whether or not they were infected with Friend leukemia virus FrC6-V, which inhibited neural differentiation at 2 x 10(4) cells/cm2. These results indicate that FrC6-V-infected P19 embryonal carcinoma cells should be seeded at high density to achieve efficient neural differentiation in vitro for ex vivo gene transfer with a FrC6-V-derived retroviral vector system.