摘要
目的比较酸性成纤维细胞生长因子(aFGF)和碱性成纤维细胞生长因子(bFGF)的生物活性。方法分离BMSCs,描绘生长曲线,进行成骨和成脂诱导鉴定;选取P3代细胞进行内皮化诱导,实验组为aFGF(10 ng/ml)+VEGF(20 ng/ml),对照组为bFGF(10 ng/ml)+VEGF(20 ng/ml),诱导第15、21天进行CD31表达率和NO分泌量的检测。结果原代细胞多为圆形和长梭形,呈集落样生长,P3代细胞均质性最佳;P1、P3代细胞生长曲线为典型的"S"型,第2～5天为对数生长期;细胞成骨、成脂诱导染色均为阳性。内皮化诱导24 d,细胞呈典型的"铺路石"样,具备内皮细胞的摄取功能;CD31表达率和NO分泌量对照组均高于实验组,统计分析显示第15天差异具有显著性,第24天差异无显著性。结论诱导BMSCs分化为ECs的过程中,aFGF与bFGF功能相同,其生物活性略低,但相对稳定,而bFGF易受细胞代谢活动的影响。
Objective To study the inducing differentiation into vascular endothelial cells （VECs） and the influence of subcuhivation on induced rate of rabbit bone marrow mesenchymal stem cells （BMSCs）. Methods Rabbit BMSCs were isolated by density gradient centrifugation methods. To identify BMSCs, the morphology of BMSCs was observed under microscope consecutively, flow cytometry was employed to detect the surface antigens including CD29, CD34, CD45 and CD90, and adipogenic and osteogenic differentiation were carried out. Vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF） were used to induce differentiation of BMSCs for 24 days. Group A and B were experimental groups, group C was control group. Cells in group A were propagated 1 generation after induced for 15 days. Cell morphogenesis, flow cytometric assay of CD31 and NO secretory volume were determined to identify the induced cells. Results The isolated cells were generally spindle shaped, attaching to the dish. The expressions of CD29, CD34, CD45 and CD90 were 98. 21% ,0. 36% ,2.03% and 86. 20%. In vitro, BMSCs were induced into osteocytes and adipocytes, Oil Red O staining and Alizarin Red S staining were positive. Induced for 24 days, BMSCs demonstrated the characters of endothelial cells, the expression of CD31 in group A, B and C were 77. 3%, 53.8% and 0. 8%, the NO secretory volume showed significant difference among the three groups. Conclusions BMSCs can differentiate into endothelial cell phenotypes induced by VEGF and bFGF in vitro, and subcuhivation can improve the induced rate.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2013年第24期6173-6175,共3页
Chinese Journal of Gerontology
基金
国家自然基金面上项目(No.30971235)
关键词
骨髓间充质干细胞
分化
血管内皮细胞
生物活性
Bone marrow mesenchymal stem cells
Differentiation
Vascular endothelial cells
Induced rate
