[The study of committed differentiation from adult rMSCs into neuron-like cells induced by Xiangdan injection in vitro].
OBJECTIVE
To study the induction of Xiangdan Injection (XDI) on the committed differentiation into neuron from rMSCs.


METHODS
The 5th passage of rMSCs were pretreated with alpha-MEM and 10 microg/L basic fibroblast growth factor (bFGF) for 24h, and treated with the serum-free induction media (D/F12 + N2 + bFGF (10 microg/L) and 1% - 5% XDI). The differentiated cells were observed with phase-contrast microscopy and detected the expression of several specific proteins, such as neurofilament-200 (NF-200), neuron-specific enolase (NSE), microtubule-associated protein-2 (MAP-2) and glial fibrillary acidic protein (GFAP) with immuno-cytochemistry and semi-quantity reverse transcriptase-polymerase chain reaction (RT-PCR).


RESULTS
At 3h of induction, rMSCs gradually turned into the neuron-like cells in the induction groups. The induction rate can reach 83.5%+/-3. 8% at 6 - 8 h of induction. The percentages of responsive cells positive for NSE,NF-200 and GFAP were 82.9%+/-2.98%, 84.1%+/-4.45% and 3.49%+/-1.67% respectively. RT-PCR showed that the relative level of expression for NSE and MAP-2 was much higher than that of GFAP (over 33 fold) (P <0. 01). The unresponsive cells in the induction group and those cultured cells in the control were shown negative for all of the for- mer detected markers.


CONCLUSIONS
The rMSCs could be differentiated into neuron-like cells in vitro with the induction of XDI in the free-serum media.
Huang
Hui
H
Neurology Department of the First Affiliated Hospital of Sun Yat-sen University, Guangzhou.
Tang
Yun'an
Y
Zhang
Cheng
C
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D005260:Female; D007150:Immunohistochemistry; D008297:Male; D009474:Neurons / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*
D000975:Antioxidants; D004365:Drugs, Chinese Herbal