中国组织工程研究与临床康复2009，Vol.13Issue(27)：5271-5275,5.DOI:10.3969/j.issn.1673-8225.2009.27.014
外源性碱性成纤维细胞生长因子对大鼠肌源性干细胞增殖的影响
Effects of exogenous basic fibroblast growth factor on proliferation of rat muscle-derived stem cells
摘要
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.关键词
肌源性干细胞/碱性成纤维细胞生长因子/浓度/培养时间分类
医药卫生引用本文复制引用
吕丹,姜殿君,张庭深..外源性碱性成纤维细胞生长因子对大鼠肌源性干细胞增殖的影响[J].中国组织工程研究与临床康复,2009,13(27):5271-5275,5.基金项目
辽宁省教委资助课题(2005L437)a grant of Education Committee of Llaoning Province,No. 2005L437 （2005L437）
