CAJ | 학술논문

目的研究在体外诱导脂肪间充质干细胞(ADMSCs)向内皮细胞分化、在立体培养基上的血管形成情况.方法将传至第三代的ADMSCs用内皮细胞诱导液、Matrigel三维培养基进行诱导培养,对ADMSCs和诱导细胞选用CD31、CD44细胞表面抗原在流式细胞仪检测表达情况,用HE染色、FⅧ-RAg免疫组织化学染色及倒置显微镜、透射电镜等进行观察鉴定.结果流式细胞仪上检测ADMSCs的表达为CD44阳性、CD31阴性,诱导的细胞CD31阳性、CD44阴性.诱导细胞14d倒置显微镜下呈铺路石样形态,FⅧ-RAg染色细胞呈阳性,透射电镜下在细胞浆内见到内皮细胞特有标志物Weibel-Palade小体。ADMSCs在Matrigel三维培养基上诱导,24h细胞迁徙成团、有伪足伸出,诱导7d伸出的细胞形成交叉网格状,13d形成较长血管,20d较长血管粗而多、出现小分叉,30d长血管、分叉血管变粗厚;FⅧ-RAg染色后诱导血管亦呈阳性.结论脂肪间充质干细胞在体外经诱导能向内皮细胞分化、能形成血管,可作为促进组织工程移植物血管化的良好的种子细胞.
목적 연구재체외유도지방간충질간세포(ADMSCs)향내피세포분화、재입체배양기상적혈관형성정황.방법 장전지제삼대적ADMSCs용내피세포유도액、Matrigel삼유배양기진행유도배양,대ADMSCs화유도세포선용CD31、CD44세포표면항원재류식세포의검측표체정황,용HE염색、FⅧ-RAg면역조직화학염색급도치현미경、투사전경등진행관찰감정.결과류식세포의상검측ADMSCs적표체위CD44양성、CD31음성,유도적세포CD31양성、CD44음성.유도세포14d도치현미경하정포로석양형태,FⅧ-RAg염색세포정양성,투사전경하재세포장내견도내피세포특유표지물Weibel-Palade소체。ADMSCs재Matrigel삼유배양기상유도,24h세포천사성단、유위족신출,유도7d신출적세포형성교차망격상,13d형성교장혈관,20d교장혈관조이다、출현소분차,30d장혈관、분차혈관변조후;FⅧ-RAg염색후유도혈관역정양성.결론 지방간충질간세포재체외경유도능향내피세포분화、능형성혈관,가작위촉진조직공정이식물혈관화적량호적충자세포.
Objective To study in vitro induction of adipose-derived mesenchymal stem cells (ADMSCs) into endothelial cells and blood vessel formation on three-dimensional (3D) media． Methods The 3rd passage ADMSCs were induced in vitro into endothelial cells on conditional medium and grew on Matrigel medium． The cell surface antigens CD31 and CD44 were detected with flow cytometric analysis before and after induction． HE staining， FⅧ-RAg immunohistochemical staining， inverted microscope and transmission electron microscope were used for morphological study.Results The expression of CD44 was positive and CD31 negative in ADMSCs in flow cytometric analysis.After induction,CD31 became positive while CD44 was negative． Paving-stone-like cell appearance was seen under inverted microscope 14 days after induction.The cells were FⅧ-RAg positively stained with immunohistological method,and Weibel-Palade body was observed under transmission electron microscope.On Matrigel media,the induced cells migrated in lumping with pseudopodia protrusion after 24 hours,and formed grid structure on the 7th day.Long vasculature was observed on the 13th day,and the vessels branched on the 20th to 30th day.The vessels stained positive for FⅧ-Rag． Conclusions ADMSCs have the potential to give rise to endothelial cells with in vitro induction and to form vessel structure in 3D media.ADMSCs have potential role in vascularized tissue engineering grafting．