摘要
目的:分离培养脂肪基质细胞(ASCs),探讨其分泌的细胞因子及其生物活性作用。方法:人脂肪组织用胶原蛋白酶Ⅰ消化后培养获得ASCs,流式细胞术鉴定其表型,ELISA法测定其分泌的血管内皮生长因子(VEGF)、肝细胞生长因子(HGF)、基质细胞衍生因子-1α(SDF-1α)等,RT-PCR测定相应mRNA表达。ASCs培养液培养人脐静脉内皮细胞,细胞计数测定内皮细胞的增殖,AnexinⅤ/PI双染色法测定内皮细胞的凋亡情况。结果:培养获得ASCs,表型为CD34+、CD105+、CD31-、CD45-,VEGF、HGF、SDF-1α的含量分别为(75±16)、(637±157)、(482±113)ng/L,能促进内皮细胞的增殖(P<0·05),降低其凋亡(P<0·05),并能上调其bcl-2的基因表达(P<0·05)。结论:ASCs能分泌细胞因子,且对内皮细胞的增殖与凋亡有明显作用。
Objective: To isolate and culture adipose stromal cells (ASCs), and study the effect of cytokines secreted by ASCs on endothelial cells. Method: Human adipose tissue was digested with collagenase type Ⅰ solution and ASCs was derived by culture. The cells surface phenotype was examined by flow cytometry. ELISA was used to detect the secretion level of VEGF, HGF,SDF-1a,and RT-PCR was used to detect the expression of their mRNA. Then the ASCs medium was used to culture human umbilical vein endothelial cells. Cells counting by hemacytometer to determined the proliferation and Annexin V / PI was used to investigate the apoptosis rate of endothelial cells. Result: ASCs was derived by culture and expressed CD34, CD105 while did not express CD31 or CD45. ASCs secreted cytokines such as VEGF, HGF and SDF-1a and the ASCs medium could stimulate proliferation and counteract apoptosis of endothelial cells ( P 〈0.05), bcl-2 mRNA was also found up regulaion in the endothelial cells. Conclusion:ASCs can secrete cytokines and have significantly effect on the endothelial cells proliferation and apoptosis.
出处
《临床心血管病杂志》
CAS
CSCD
北大核心
2006年第10期624-627,共4页
Journal of Clinical Cardiology
基金
湖北省科技攻关计划(No:2005AA304B03)
关键词
脂肪基质细胞
细胞因子
内皮细胞
Adipose stromal cells
Cytokines
Endothelial cells