The present work deals with the finding and characterization of a neurotrophic factor present in serum-free Dulbecco's modified Eagle's medium in which rat sciatic nerves previously cultured for 9 days were maintained for 24 h. This sciatic nerve conditioned medium (SNCM) produced neuronal differentiation and neurite outgrowth on PC12 cells, as well as survival and differentiation of eight-day old chick embryo dorsal root ganglion (E8-DRG) and ciliary ganglion (E8-CG) neurons. SNCM activity was decreased by dilution, heating and trypsin treatment; it was not inhibited by anti-NGF and anti-bFGF antibodies; and it was not mimicked by CNTF, laminin and fibronectin. By utilizing its neurite-promoting activity on PC12 cells, experiments oriented to purify the factor were carried out. Ultrafiltration, heparin-affinity chromatography and size-exclusion high pressure liquid chromatography (HPLC) were employed. The ability of SNCM to induce PC12 cell, E8-DRG and E8-CG neuronal differentiation, the heparin affinity of the active SNCM protein, and the size-exclusion HPLC elution characteristics of the active protein suggest that the active component of the SNCM is, in all probability, a novel sciatic nerve neurotrophic factor (SNTF).
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