大鼠心脏周细胞的提取、培养及鉴定
首发时间:2016-05-25
摘要:目的:建立简便、高效、稳定的大鼠心脏周细胞的原代提取及培养方法。方法:健康雄性SD大鼠,经酶消化、过滤、离心后进行心脏周细胞的原代培养;倒置相差显微镜观察细胞生长情况,苏木素-伊红染色方法观察细胞形态;免疫荧光进行细胞鉴定;Western Blot定量细胞鉴定。结果:倒置相差显微镜下原代细胞接种后24小时细胞呈梭型或星芒状,3-5天可达到95%左右融合状态。HE染色显示,周细胞呈不规则多边形,胞质丰富,嗜酸性,胞核位于胞体中央,圆形或长椭圆形,偶见双核,部分细胞排列紊乱,交叉重叠生长,无接触性抑制。免疫荧光染色观察到PDGFR-β、NG2 、CD146、COL4A5阳性,CD146和PDGFR-β双阳性及NG2和PDGFR-B双阳性,CD31阴性的贴壁细胞;Western Blot定量分析CD146、PDGFR-β、NG2 有表达,CD31无表达。结论:成功的建立高效、简单心脏周细胞原代提取的方法。
For information in English, please click here
Isolation、Culture and Identify of Pericytes from Rat Heart
Abstract:Objective: To establish a simple, efficient and stable method for the isolation of rat heart pericytes in vitro. Methods: Primary cardiac pericytes were obtained after enzyme digestion, filtration, and centrifugation for healthy male SD rats. Inverted microscope was used to observe the cell growth and cell morphology after hematoxylin-eosin staining. Immunofluorescence staining was used to identify the cells. Western blot was used to quantitatively cell identification. Results: Primary cardiac pericytes were spindle or star in 24 hours after inoculation, and the fusion status was about 95% in 3-5 days. HE staining showed that, primary cardiac pericytes showed irregular polygon, abundant cytoplasm, eosinophilic nuclei in the central cell body, round or oval and occasionally binucleate, cells arranged in disorder, overlapping growth without contact inhibition. Immunofluorescence staining was observed in PDGFR-β, NG2, CD146, COL4A5+, CD146 and PDGFR-β double positive and NG2 and PDGFR-β double positive and negative for CD31 adherent cells. Western blot quantitative analysis of CD146, PDGFR-β, NG2 expression of CD31 expression. Conclusion: Method of high efficiency and simple extraction of primary cardiac cycle cells was successfully established.
Keywords: Pericytes Heart Primary cell extraction
论文图表:
引用
No.4692075114907014****
同行评议
勘误表
大鼠心脏周细胞的提取、培养及鉴定
评论
全部评论0/1000