Research of autophagy activity between rat bone marrow mesenchymal stem neural differentiation.
- Author:
Bo LI
;
Chun-tian HUANG
;
Cai-fang LI
;
Ping DUAN
;
Xue-fei HAN
;
Wen-hai YAN
;
Ying XING
- Publication Type:Journal Article
- MeSH: Animals; Autophagy; Cell Differentiation; Cells, Cultured; Flow Cytometry; Mesenchymal Stromal Cells; cytology; Neurons; cytology; Rats
- From: Chinese Journal of Applied Physiology 2015;31(1):31-34
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.
METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.
RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.
CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.