[Effect of different FSB concentration and first-time exchange of total volume medium on proliferation of bone marrow stromal cells from GFP transgenic mouse in vitro].
OBJECTIVE
To evaluate the effect of different fetal bovine serum (FBS) concentration and first-time exchange of total volume medium on the proliferation of bone marrow stromal cells (BMSCs) from green fluorescence protein (GFP) transgenic mouse in vitro.


METHODS
Bone marrow cells isolated from GFP transgenic mice were cultured in DMEM/F12 containing 10%, 20%, 30% FBS respectively; the first exchange of the total volume medium was made at different times (4 h, 6 h, 8 h, 10 h, 12 h, 24 h, 48 h and 72 h) after 3 d primary culture; then the total volume medicum was exchanged every three days. The amplification of BMSCs was determined. The passage 5 BMSCs cultured in DMEM/F12 containing 10% and 20% FBS were examined with the antibodies CD44, CD45 and CD54 at the time of first exchange of the total volume medium.


RESULTS
The cultured cells proliferated well in DMEM/F12 containing 10% FBS and 20% FBS. With the extension of the time for first exchange of total volume medium, the density of the adhered cells increased, but the purity of BMSCs decreased.


CONCLUSIONS
The method of making cells adhere to culture plastic in different time for cultivating and purifying BMSCs from GFP transgenic mice is effective, the appropriate concentration of FBS is 10%-20% and the best time for the the first exchange of total volume medium is 8 hour.
Shi
Zhen-jiang
ZJ
The Institute of Neuroscience, Kunming Medical College, Kunming 650031, China.
Huang
Gui-qin
GQ
Wang
Ting-hua
TH
Xu
Wei
W
Li
Fu-qiang
FQ
Feng
Zhong-tang
ZT
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002455:Cell Division; D002478:Cells, Cultured; D003470:Culture Media; D049452:Green Fluorescent Proteins / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D011897:Random Allocation; D044967:Serum; D017154:Stromal Cells / Q000166:cytology*
D003470:Culture Media; D049452:Green Fluorescent Proteins