bFGF and heparin but not laminin are necessary factors in the mediums that affect NSCs differentiation into cholinergic neurons.
OBJECTIVE
Neural stem cells (NSCs) are self-renewed, pluripotent cells that can differentiate into neurons, astrocytes and oligodendrocytes. Cholinergic neurons are an important kind of neurons that play an essential role in the treatment of Parkinsonism and epilepsy. We are interested in how different mediums affect NSCs differentiation into cholinergic neurons.


METHODS
NSCs were isolated from the striatum corpora of embryonic brain in a 14-day pregnant rat. Cells were cultured in basic mediums [F12/DMEM (1:1) including 1% B27 (v/v) and 20 ng/ml EGF] but with different combinations of three supplements: bFGF (20 ng/ml), heparin (5 mug/ml) and laminin (1 mug/ml). After 7 days culturing, cells were immunized with choline acetyltransferase (ChAT), a marker enzyme of cholinergic neuron.


RESULTS
We found ChAT could not be detected in the basic mediums with only one supplement. Then, we tested the combination of two out of three. We found that ChAT positive cells could only be detected in the medium with bFGF and heparin (FH). However, when we added the laminin into the FH, more ChAT positive cells appeared.


CONCLUSIONS
This finding suggests that bFGF and heparin are essential in the mediums that affect NSCs differentiation into cholinergic neurons, and laminin is an important positive factor in this process.
Ren
Wenwen
W
School of Life Sciences, Shanghai University, 99 Shangda Road, Shanghai 200444, China.
Guo
Qigen
Q
Yang
Yang
Y
Chen
Fuxue
F
D000818:Animals; D015152:Blotting, Northern / Q000379:methods; D001921:Brain / Q000166:cytology; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism*; D004622:Embryo, Mammalian; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000502:physiology*; D006493:Heparin / Q000502:physiology*; D007150:Immunohistochemistry / Q000379:methods; D007381:Intermediate Filament Proteins / Q000378:metabolism; D007797:Laminin / Q000502:physiology; D008297:Male; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D013234:Stem Cells / Q000166:cytology*
D007381:Intermediate Filament Proteins; D007797:Laminin; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D006493:Heparin; D002795:Choline O-Acetyltransferase