小鼠胚胎干细胞体外分化形成新生血管的实验研究  

AN EXPERIMENTAL STUDY OF NEW BLOOD VESSEL FORMATION FROM IN VITRO DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELLS

作　　者：韩雅玲[1] 田孝祥[1] 康建[1] 徐凯[1] 闫承慧[1] 

机构地区：[1]沈阳军区总医院全军心血管病研究所心内科,沈阳110016

出　　处：《中国组织化学与细胞化学杂志》2006年第6期592-597,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金资助项目(30370526)

摘　　要：目的探讨小鼠胚胎干细胞(ES)在体外向内皮细胞(ECs)分化并形成新生血管的条件及特点。方法分别建立二维和三维小鼠ES细胞分化体系,对分化细胞行血小板内皮细胞粘附分子(PECAM-1)免疫荧光染色和DiI-乙酰化低密度脂蛋白(DiI-Ac-LDL)标记,观察ECs分化及血管形成特点。结果在二维分化体系,ES细胞在无外源性生长因子存在的条件下可自发向ECs分化,ECs主要定位在分化细胞密集处,分化表现为:ECs呈集落样生长,不形成网状结构;ECs互相连接形成网状结构,PECAM-1免疫荧光染色证实为ECs网。在三维的悬浮拟胚体培养体系,ECs的分化不依赖于外源性生长因子,分化表现为条索状结构、管腔样结构及排列紊乱的细胞团三种形式。对该拟胚体的冷冻切片进行的三维图像重构显示,拟胚体中有大量的血管网形成。在三维的Ⅰ型胶原培养体系中,ES形成的拟胚体表现为出芽式血管新生,这一过程依赖于外源性生长因子混合物。结论在二维和三维ES细胞分化体系中,ECs分化及新生血管形成过程与体内相似,表现为血管生成和血管新生两种形式,因此可作为研究血管发育机制的理想模型。Objective To explore the conditions and characteristics of endothelial cell （EC） differen tiation as well as new blood vessel formation from in vitro differentiation of mouse embryonic stem （ES） cells. Methods Two-dimensional and three-dimensional differentiation systems of mouse ES cells were established respectively, and then platelet-endothelial cell adhesion molecule-1 （PECAM-1） immunofluorescent staining and DiI-acetylated low-density lipoprotein （DiI-Ac-LDL） labeling of differentiated cells were performed to observe the characteristics of EC differentiation and blood vessel formation. Results In the two-dimensional differentiation system, ES cells differentiated into ECs in the absence of exogenous growth factors. The ECs were localized in areas with high cell density. Some differentiated ECs grew as a colony instead of endothelial networks, others connected to form EC networks confirmed by PECAM-1 immunofluorescent staining. In the suspended embryoid bodies （EBs） culture system of three-dimensional differentiation, three structures were found without addition of exogenous growth factors, cord-like structure, vessel-like structure and disordered cell mass. The three-dimensional reconstruction of these EBs＂ cryosections showed massive vascular networks. In the type Ⅰ collagen culture system of three-dimensional differentiation, EBs derived from ES cells displayed sprouting angiogenesis, which was dependent on cocktails of exogenous growth factors. Conclusion In these two-and three-dimensional differentiation systems in vitro, EC differentiation and blood vessel formation are similar to the in vivo process, which includes vasculogenesis and angiogenesis. Thus these two systems may serve as ideal models for further research on mechanisms of vascular development.

关 键 词：胚胎干细胞 内皮细胞 血管生成 血管新生 分化 

分 类 号：Q954.4[生物学—动物学]