摘要
目的 观察脂肪来源的间充质干细胞（ad-MSCs）在经过传代扩增后诱导内皮细胞方向的分化能力.方法 使用胶原酶消化法分离培养原代脂肪间充质干细胞,检测其生长特性并用流式细胞术对其进行了联合免疫表型鉴定,诱导其多向分化来确定其干细胞特性.扩增后的高代次（P5）脂肪间充质干细胞被用于进行3种条件下的诱导内皮细胞方向分化（基础培养基＋血管内皮生长因子（VEGF）、内皮细胞支持液＋VEGF、仅用内皮细胞支持液）,诱导时间为12 d,之后用流式细胞术检测CD31并用免疫细胞化学法检测Ⅷ因子表达来评估分化效果.结果 我们成功地进行了脂肪间充质干细胞的原代培养,原代培养的细胞表现出了成纤维细胞样形态,在免疫表型和多向分化能力都表现出了干细胞特性.经过仅12 d的诱导内皮细胞分化后,EGM2-MV＋ VEGF组开始表达CD31和Ⅷ因子,而另外两组未发现表达.结论 传代扩增后的高代次的脂肪间充质干细胞仍有较强的内皮细胞分化能力,EGM2-MV＋ VEGF能更高效地促进其向内皮细胞分化.
Objective To research amplified high generation human adipose-derived mesenchymal stem cells （ad-MSCs） endothelial differentiated capacity.Methods Ad-MSCs were isolated by collagenase I and then cultured and amplified,analyzed the surface markers and tested the multilineage differentiated potential to demonstrate stem cells＇ characteristics,passage 5 cells were specially selected to study their endothelial differentiated capacity.Three different induced conditions （low-serum basic medium ＋ 50 μg/L vascular endothelial growth factor （VEGF）,EGM2-MV ＋ 50 μg/L VEGF and single EGM2-MV） were designed to explore an optimal method to achieve the highest differentiated efficiency,and then the differentiated efficiency was evaluated by CD31 and Ⅷ factor expression.Results Flow cytometry surface markers analysis and multilineage differentiation tests demonstrated the cultured cells stem cells characteristics.In the process of differentiating into endothelial cells,passage 5 ad-MSCs in EGM2-MV ＋ VEGF group acquired endothelial markers CD31 and partly expressed Ⅷ factor after only 12 days inducing.Conclusion Amplified high generation ad-MSCs still have considerable differentiated potential towards endothelial cells,and EGM2-MV containing high concentrations of VEGF can be more effective for endothelial differentiation than two other groups.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第11期2405-2407,共3页
Chinese Journal of Experimental Surgery
基金
中国博士后科学基金资助项目（20100481271）
山东省自然科学基金资助项目（ZR2012HM002）
山东大学自主创新基金资助项目（2011JC016）
关键词
间充质干细胞
内皮细胞
分化
组织工程
Mesenchymal stem cells
Endothelial cells
Differentiation
Tissue engineering
