Human umbilical cord mesenchymal stem cells differentiate into neuron-like cells after induction with B27-supplemented serum-free medium.
10.12122/j.issn.1673-4254.2020.09.18
- Author:
Yunyi LI
1
;
Jinpei YANG
1
;
Guo FU
1
;
Pan ZHOU
1
;
Yang LIU
1
;
Zhizhong LI
2
;
Genlong JIAO
1
Author Information
1. First Affiliated Hospital of Jinan University, Guangzhou 510632, China.
2. Shenhe Hospital Affiliated to Jinan University, Heyuan 517000, China.
- Publication Type:Journal Article
- Keywords:
differentiation;
human umbilical cord mesenchymal stem cells;
neuron-like cells;
serum-free medium
- From:
Journal of Southern Medical University
2020;40(9):1340-1345
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To evaluate the capacity and efficiency of human umbilical cord mesenchymal stem cells (HUCMSCs) to differentiate into neuron- like cells after induction with B27- supplemented serum- free medium.
METHODS:HUCMSCs at passage 4 were cultured for 14 days with serum-containing medium (SCM) (group A), SCM supplemented with 20 ng/mL nerve growth factor (NGF) and 10 ng/mL basic fibroblast growth factor (bFGF) (group B), serum-free medium (SFM) (group C), or SFM supplemented with 20 ng/mL NGF and 10 ng/mL bFGF. The culture medium were changed every 3 days and the growth of the neurospheres was observed using an inverted microscope. The cell markers were analyzed with flow cytometry and the expressions of nestin, neuron- specific enolase (NSE), neurofilament heavy polypeptide (NEFH), and glial fibrillary acidic protein (GFAP) were quantified by quantitative real-time PCR (qRT-PCR) and Western blotting.
RESULTS:Before induction, HUCMSCs expressed abundant mesenchymal stem cell surface markers including CD29 (99.5%), CD44 (49.6%) and CD105 (77.7%). Neuron-like cells were observed in the cultures on days 7, 10, and 14, and the cell differentiation was the best in group D, followed by groups C, B and A. In all the 4 groups, the cellular expressions of nestin and GFAP gradually lowered while those of NEFH and NSE increased progressively. The expressions of GFAP, NEFH, nestin and NSE were significantly different between group A and the other 3 groups ( < 0.001 or 0.05).
CONCLUSIONS:B27-supplemented SFM effectively induces the differentiation of HUCMSCs into neuron- like cells, and the supplementation with cytokines (NGF and bFGF) strongly promotes the cell differentiation.
