[Extraction and purification of Schwann cells cytoplasmic neurotrophic proteins with high pressure liquid chromatography and study on its neurobiological activities].
OBJECTIVE
To purify and study Schwann cells cytoplasmic neurotrophic protein.


METHODS
The dissociated SC taken from 300 newborn rats sciatic nerves were cultured, collected, ultrasonicated and ultraspeed centrifuged. The supernates were ultrafiltrated and concentrated by using ultrafiltration units with PM10, PM30, PM50 ultrafiltration membranes. The ultrafiltrated-concentrated solution with the protein molecular weight 10-30 ku, 30-50 ku and > 50 ku were collected respectively. The dissociated spinal cord motoneurons of 14 days embryonic rats were cultured with serum-free conditional medium and the additional SC cytoplasmic proteins were added into the medium. The results showed that the 10-30 ku and > 50 ku SC cytoplasmic proteins were able to maintain the survival of motoneurons for 24 hours. Then the 26 ku and 58 ku proteins were further extracted and purified from SC cytoplasm by high pressure liquid chromatography, and their neurobiological activities were studied.


RESULTS
The 26 ku and 58 ku Schwann cell's cytoplasmic proteins were able to maintain the survival of motoneurons cultured in the serum-free medium for 48 hours. The highest biological activity concentration is 20 ng per well.


CONCLUSIONS
Schwann cells cytoplasm contains motoneuron neurotrophic proteins with molecular weight 26 ku and 58 ku.
Xie
G
G
Department of Plastic and Microsurgery, First Affiliated Hospital, Sun Yat-sen University of Medical Sciences, Guangzhou, Guangdong, P. R. China 510080.
Zhu
J K
JK
Gu
X F
XF
D000818:Animals; D000831:Animals, Newborn; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003593:Cytoplasm / Q000737:chemistry; D009046:Motor Neurons / Q000166:cytology; D009414:Nerve Growth Factors / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000737:chemistry* / Q000166:cytology; D012584:Sciatic Nerve / Q000737:chemistry* / Q000166:cytology
D009414:Nerve Growth Factors