摘要
目的:介绍大鼠乳鼠心房肌细胞的分离、纯化,以及培养和鉴定方法。方法:取1 d龄SD大鼠心房组织,用0.1%胰蛋白酶和0.025%Ⅱ型胶原酶消化,将心房肌细胞收集在含10%胎牛血清的DMEM培养基中,用差速贴壁和5-溴脱氧尿嘧啶核苷纯化心房肌细胞。观察细胞形态,结合α-横纹肌肌动蛋白抗体免疫荧光鉴定心房肌细胞。结果:细胞培养24 h已完全贴壁,成梭形或三角形,无细胞搏动,细胞体积可逐渐增大。培养细胞经免疫荧光鉴定,95%为心房肌细胞结论:该研究是一种较好的乳鼠心房肌细胞原代培养及鉴定方法。
Objective：To introduce a method for isolation, purification, culture and identification of the atrial myocytes of neonate rat. Methods：After digested by trypsin （0.1%） and type Ⅱ collagenase （0. 025%）, the atrial myocytes were isolated from atrial tissue of the Sprague Dawley rats one day old and were collected in DMEM medium with 10%fetal bovine serum, then were purified with the differential adhesion method and 5-bromodeoxyuridine process method. Atrial myocytes were identified by cell morphology combined with a-striated muscle actin antibody immunofluorescence staining. Results：After 24 hours, the cultured cells completely adhered and were in triangle or spindle-shape without beating. The cell volume was gradually increased. With immunofluorescence staining, 95 % cells were identified to be atrial myocytes. Conclusion： Our method is suitable for primary culture and identification of the atrial myocytes of neonate rats.
出处
《国际心血管病杂志》
2013年第2期112-115,共4页
International Journal of Cardiovascular Disease
基金
国家自然科学基金(81060018)
关键词
乳鼠
细胞培养
心房肌细胞
Neonate rat
Cell culture
Atrial myocytes
