摘要
目的 研究不同浓度TGF-β1对大鼠骨髓内皮祖细胞（endothelial progenitor cells，EPCs）体外增殖的影响。优化EPCs体外培养条件。方法 冲洗大鼠骨髓腔，密度梯度离心得到单个核细胞；通过fibronectin包被的培养皿差速分选、富集EPCs，利用流式细胞仪对所富集细胞进行CD34，CD133，VEGFR-2表型鉴定。将EPCs在含有10～300pg／ml的不同浓度TGF-β1的M199培养基中传代培养扩增，记录细胞扩增数量，并应用流式细胞仪对不同代次细胞，进行CD34，CD133和VEGFR-2表达检测。结果 快速黏附于fibronectin的细胞群中，CD34，CD133，VEGFR-2阳性细胞含量明显高于未分选细胞（P〈0．05）。细胞培养至P4，在TGF-β1浓度为10pg／ml的实验组，细胞总体扩增数量以及CD34，CD133，VEGFR-2阳性细胞含量，均明显高于其他浓度TGF-β1的实验组（P〈0．05）。结论 培养基中低浓度的TGF-β1能提高大鼠骨髓EPCs体外扩增数量，而且减慢了EPCs的分化速度。从而使EPCs发挥更高的治疗效率。
Objective To investigate the proliferation of rat bone marrow endothelial progenitor ceils （EPCs） in the culture under different concentrations of TGF-β1 in order to optimize the culture conditions. Methods EPCs harvested from bone marrow by flushing fresh rat femur and tibia were isolated by density gradient eentrifugation. The isolated cells were further purified and enriched by fast adhere to fibroneetin coated dish. Flow eytometry was applied to enrieh the cells positive to CD34, CD133 and VEGFR-2. EPCs were expanded in M199 medium in presence of different concentrations of TGF-β1 （ 10 -300 pg/ml）. Total eell output was recorded; and the expressions of CD34, CD133, and VEGFR-2 at different cell passages were analyzed through flow eytometer. Results Fast adhere cell group showed significantly higher positive proportion of CD34, CD133, and VEGFR-2 than unsorted cells （P 〈0.05）. At passage 4, in the presence TGF-bl at low concentration （10 pg/ml） euhured EPCs demonstrated significantly higher positive proportion of CD34, CD133, VEGFR-2 than control groups （P 〈0.05）. Conclusion TGF-β1 at low concentrations （10 pg/ml） induces a very significant additional increase of expansion of rat EPCs as well as slows down the differentiated speed, which promotes their therapeutie application.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2006年第23期2306-2308,共3页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目(30570517)~~
关键词
TGF-Β1
EPC
体外
增殖
TGF-β1
endothelial progenitor cells
proliferation
