CAJ | 학술논문

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.
목적 관찰지방래원적간충질간세포(ad-MSCs)재경과전대확증후유도내피세포방향적분화능력.방법 사용효원매소화법분리배양원대지방간충질간세포,검측기생장특성병용류식세포술대기진행료연합면역표형감정,유도기다향분화래학정기간세포특성.확증후적고대차(P5)지방간충질간세포피용우진행3충조건하적유도내피세포방향분화(기출배양기+혈관내피생장인자(VEGF)、내피세포지지액+VEGF、부용내피세포지지액),유도시간위12 d,지후용류식세포술검측CD31병용면역세포화학법검측Ⅷ인자표체래평고분화효과.결과 아문성공지진행료지방간충질간세포적원대배양,원대배양적세포표현출료성섬유세포양형태,재면역표형화다향분화능력도표현출료간세포특성.경과부12 d적유도내피세포분화후,EGM2-MV+ VEGF조개시표체CD31화Ⅷ인자,이령외량조미발현표체.결론 전대확증후적고대차적지방간충질간세포잉유교강적내피세포분화능력,EGM2-MV+ VEGF능경고효지촉진기향내피세포분화.
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.