Neuronal differentiation of PC12 and chick embryo ganglion cells induced by a sciatic nerve conditioned medium: characterization of the neurotrophic activity.
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).
Villegas
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Instituto Internacional de Estudios Avanzados (IDEA), Apartado, Caracas, Venezuela.
Haustein
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Villegas
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D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002642:Chick Embryo; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D005260:Female; D005726:Ganglia, Parasympathetic / Q000166:cytology* / Q000196:embryology; D005727:Ganglia, Spinal / Q000166:cytology* / Q000196:embryology; D008297:Male; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009474:Neurons / Q000166:cytology*; D016716:PC12 Cells; D051381:Rats; D012584:Sciatic Nerve / Q000502:physiology*; D014357:Trypsin; D014462:Ultrafiltration
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D009414:Nerve Growth Factors; D014357:Trypsin