体外诱导脂肪干细胞向内皮细胞及成血管的分化  

In vitro induction of adipose-derived stem cells into endothelial cells and angiogenic cells

作　　者：马丽[1] 黄鲁刚[1] 林苹[2] 

机构地区：[1]四川大学华西医院小儿外科,四川省成都市610041 [2]四川大学华西医院老年医学研究室,四川省成都市610041

出　　处：《中国组织工程研究》2013年第10期1730-1735,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:脂肪干细胞作为组织工程中种子细胞的选择,可以诱导为内皮细胞从而有效解决材料血管化困难的难题。目的:体外观察脂肪干细胞经诱导向内皮细胞转分化的可能性、以及在立体培养基上的血管形成情况。方法:切取人皮下脂肪,用酶消化法分离和培养脂肪干细胞,将传至第3代或第4代的脂肪干细胞用内皮细胞诱导液、同时在Matrigel三维培养基内诱导培养,观察细胞生长情况及变化。对脂肪干细胞和诱导细胞行免疫组织化学检查CD31的表达。结果与结论:免疫组织化学检测脂肪干细胞的CD31表达阴性,诱导细胞CD31可见阳性表达。在三维立体培养基内诱导的细胞24h逐渐迁徙成团,伸出伪足,诱导1周细胞形成网格样交叉,2周形成较长血管,后血管增粗,并出现分叉,CD31阳性表达。因此,脂肪干细胞体外可以被诱导向内皮细胞表型转分化,并形成血管,提示脂肪干细胞可以作为促进组织工程移植物血管化的种子细胞的理想选择。BACKGROUND： Adipose-derived stem cells, as the seed cells for tissue engineering, can be induced into endothelial cells, thereby solving the key problem of vascularization. OBJECTIVE： To in vitro investigate the feasibility of inducing adipose-derived stem cells to differentiate into endothelial cells and the blood vessel formation on three-dimensional media. METHODS： Adipose-derived stem cells were isolated and cultured by enzyme digestion. Passage 3 or 4 adipose-derived stem cells were induced into endothelial cells with conditional medium and then the induced cells were cultured in Matrigel medium. Cell surface antigen CD31 was detected by immunohistochemistry before and after induction. After hematoxylin-eosin staining, cell morphology was observed by phase contrast microscopy.RESULTS AND CONCLUSIONS： Immunohistochemical examination results showed that CD31 expression was negative in adipose-derived stem cells, but positive expression was detected in the induced cells. The cells cultured in Matrigel media accumulated together gradually, emanated pseudopodia after 24 hours and formed grid structure after 1 week, and long vasculature was observed after 2 weeks. CD31 expression was detected in the induced cells. These results indicate that adipose-derived stem cells can be in vitro induced into endothelial cells, then which form vasculature, suggesting that adipose-derived stem cells can be used as the ideal seed cells for vascularization in tissue-engineered grafts.

关 键 词：干细胞 脂肪干细胞 内皮细胞 血管 诱导 CD31 三维立体培养 细胞培养 脂肪组织 分化 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]