[Extraction and purification of neurotrophic proteins from bone marrow stromal cells].
OBJECTIVE
To extract and purify the cytoplasmic neurotrophic proteins from bone marrow stromal cell and to test their neurotrophic activity.


METHODS
Bone marrow stromal cells were collected from rat femur, after ultrasonic grind and ultracentrifugation, the supernate was ultrafiltered and concentrated, the proteins that molecular weight was greater than 10 ku were collected. The neurotrophic active substance was extracted and purified by Sephadex G-100 gel chromatography and high performance liquid chromatography (HPLC). Then their neurotrophic activity was tested in cultured spinal cord motoneuron with MTT method and morphologic observation.


RESULTS
After supernate was analyzed by Sephadex G-100 chromatography, peak I could promote the growth of neuron. A further analysis of peak II with HPLC showed that peak A could promote the growth of neuron. The SDS-PAGE analysis of peak A indicated that a main protein zone with molecular weight of 13 ku was obtained.


CONCLUSIONS
The protein of 13 ku in MSCs has neurotrophic activity.
Zhou
Liyi
L
Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei, 430022, PR China.
Hong
Guangxiang
G
Wang
Fabin
F
D000818:Animals; D001854:Bone Marrow Cells / Q000737:chemistry*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D009046:Motor Neurons / Q000166:cytology / Q000187:drug effects; D009414:Nerve Growth Factors / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D013116:Spinal Cord / Q000166:cytology; D017154:Stromal Cells / Q000737:chemistry*
D009414:Nerve Growth Factors