血管类器官的构建及鉴定  

Generation and identification of blood vessel organoids

作　　者：刘琪 赵海静 金蕊 程龙 刘昱圻 Liu Qi;Zhao Haijing;Jin Rui;Cheng Long;Liu Yuqi(Chinese PLA Medical School,Department of Cardiology,Chinese PLA General Hospital No.1 Medical Center,Beijing 100853,China)

机构地区：[1]解放军医学院解放军总医院第一医学中心心血管内科,100853 [2]军事科学院军事医学研究院生物工程研究所

出　　处：《中华老年心脑血管病杂志》2022年第5期527-530,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(82070434);北京市科技新星交叉项目资助(Z191100001119020)。

摘　　要：目的 探讨人诱导多能干细胞(hiPSC)培养血管类器官的方法并进行鉴定。方法 体外培养hiPSC,先诱导hiPSC形成拟胚体,再诱导分化中胚层和血管祖细胞后,在Ⅰ型胶原蛋白-基质胶介质中分化形成血管网,分离单个血管网,形成血管类器官;倒置显微镜观察分化过程中拟胚体,血管网及血管类器官的生长情况;免疫荧光染色检测血管类器官的内皮细胞特异性抗体CD31,平滑肌细胞特异性抗体α平滑肌肌动蛋白(α-SMA),血小板衍生生长因子受体β(PDGF-β)蛋白表达。结果 构建血管类器官3D细胞模型,免疫荧光染色显示,CD31阳性,α-SMA阳性,PDGF-β阳性,与人体血管组织结构特征相似。结论 初步构建了血管类器官3D细胞模型,将为心血管疾病的基础研究和靶向药物的筛选提供新的模型。Objective To study how to culture and identify blood vessel organoids from human induced pluripotent stem cells(hiPSC).Methods The embryoid bodies were developed by inducing the in vitro cultured hiPSC,then the vascular nets were formed in type Ⅰ collagen matrigel medium by inducing and differentiating the mesoderm and vascular progenitor cells, the blood vessels organoids were finally generated by isolating individual vascular net.The growth of differentiating embryoid bodies, vascular nets and blood vessel organoids was observed by inverted microscopy.The expressions of endothelial cell specific antibody CD31,smooth muscle cell specific antibody α(α-SMA),and platelet-derived growth factor receptor β(PDGF-β) protein in blood vessel organoids were detected with immunofluorescence staining.Results The 3 D cell model of blood vessel organoids was established.Immunofluorescence staining showed positive CD31,α-SMA,and PDGF-β,which were similar to the structural characteristics of human blood vessels.Conclusion The 3 D cell model of blood vessel organoids established in this study can provide a new model for basic research of cardiovascular disease and screening of targeted drugs.

关 键 词：干细胞 器官培养技术 人工器官 血管 

分 类 号：R-33[医药卫生]