Proliferating and differentiating Schwann cell cultures from embryonic chick sciatic nerve maintained for months in vitro without antimitotics or growth factors.

In general, the available methods for culturing Schwann cells require specific antibodies and/or the addition of antimitotics to suppress fibroblasts, plus various factors to support their growth. Moreover, the maximal culture period of Schwann cells normally is limited to a few weeks. Here, three easy novel methods to culture Schwann cells from embryonic chick sciatic nerve are presented, that require no growth factors or agents elevating intracellular cAMP. In contrast to the conventional antimitotic treatment with cytosine arabinoside, we use D-valine to suppress fibroblasts. Our modified medium C leads within a few days to highly enriched Schwann cell cultures (culture I). Passage into a serum-reduced medium D allows for differentiating longterm cultures (culture II). In cultures I and II, the rate of cell division is low. However, after passage into serum-containing SC-medium, proliferation increases within one week to high levels (culture III). Cultures II and III can be grown for several months, during which time spontaneous immortalization can occur. The high purity of the cultures of about 95% is assessed using glia-specific antibodies for S-100 antigen, HNK-1 epitope, glia fibrillary acidic protein (GFAP), galactocerebroside (Gal C) and 3A7. These culture procedures are easy to perform and are suitable for differentiation, proliferation and coculturing experiments.