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.
Conclusion: 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.