Effects of exogenous basic fibroblast growth factor on proliferation of rat muscle-derived stem cells

Dan LU; Dianjun JIANG; Tingshen ZHANG

ABSTRACT

BACKGROUND:

Muscle-derived stem cells have some problems in pudflcation, amplification and directional differentiation during in vitro culture. Basic fibroblast growth factor as a membrane of bloactive factor has been paid great attention, due to its biological activity.

OBJECTIVE:

To explore effects of basic flbroblast growth factor on/n vitro proliferation of rat musde-derived stem cells.DESIGN, TIME AND

SETTING:

The cytological in vitro study was performed at the Shenyang Medical College from April to July 2008.MATERIALS Five healthy male Wistar rats were supplied by Experimental Animal Center, Shenyang Medical College. Basic fibroblast growth factor was produced by Zhuhai Biological Product Co., Ltd.

METHODS:

Left anterior limb triceps brachii was sterilely obtained from rats. Muscle-derived stem cells were harvested by isolation with enzyme digestion, pudfied by density gradient centrifugation and differential attachment. At the second passage,musde-derived stem cells were treated with basic fibroblast growth factor at final concentration of 6.25, 12.50, 25.00, 50.00,100.00 pg/L. Cells received 200 uL growth medium as blank control group, and those treated with 200 μL growth medium +musde-derived stem celts as negative controls. MTT was added at 24, 48, 72 and 96 hours of culture.MAIN OUTCOME

MEASURES:

Immunocytochomical method was used to determine rat musde-dedved stem cells. MTT assay was utilized to detect cell proliferation.

RESULTS:

Musde-denved stem cells were mostly positive for Sca-1, but few was positive for Desmin. Compared with the negative control group, vanous concentrations of basic fibroblast growth factor had significant proliferation effects on musde-derived cells (P < 0.05). At 6.25-50.00 μg/L, the proliferation effect became strong with the increased concentration (P <0.01). At 50.00 μg/L, the proliferation promotion effect nearly reached a peak. Musde-derived stem cells significantly proliferated over time. Compared with the negative control group, significant proliferation promotion effect was detected in the basic fibroblast growth factor groups at 96 hours (P < 0.01).

CONCLUSION:

Basic fibroblast growth factor can promote proliferation of rat muscle-derived stem cells/n vitro in a dose-time dependent fashion. 50.00 μ g/L and 96 hours are separately good concentration and time for in vitro culture.