Smith D A, Glover J L, Townsend L E, Maupin D E
William Beaumont Hospital, Royal Oak, Michigan 48073.
In Vitro Cell Dev Biol. 1991 Dec;27A(12):914-20. doi: 10.1007/BF02631117.
Myocardial cell culture methods are now well established for animal and fetal human tissue. We present here a method for harvesting and culturing adult human atrial myocardiocytes. Cells are obtained from fresh atrial tissue normally discarded after being removed to cannulate the right atrium during open heart surgery. The atrial tissue is minced and then digested using collagenase. The single cell suspension is initially cultured in serum-containing growth medium, then transferred to defined medium, selective for myocardial cell growth. The cells are characterized by immunoperoxidase stains and transmission electron microscopy. The cultured cells stain positive for myoglobin, whereas control cultured fibroblasts and endothelial cells do not. Electron microscopy shows the presence of numerous myofibrils, Z-bodies, pleomorphic mitochondria, and secretory granules. The chronological age of the donor was an important factor in culturing the adult tissue, the younger tissue correlated with a higher success rate. This method provides a means for in vitro study of human adult myocardial cells and provides guidelines for appropriate atrial tissue to use.
心肌细胞培养方法目前在动物和胎儿人体组织中已得到充分确立。我们在此介绍一种获取和培养成人心房心肌细胞的方法。细胞取自心脏直视手术中为插管进入右心房而切除的通常会被丢弃的新鲜心房组织。将心房组织切碎,然后用胶原酶消化。单细胞悬液最初在含血清的生长培养基中培养,然后转移至对心肌细胞生长有选择性的限定培养基中。通过免疫过氧化物酶染色和透射电子显微镜对细胞进行鉴定。培养的细胞肌红蛋白染色呈阳性,而对照培养的成纤维细胞和内皮细胞则无此现象。电子显微镜显示存在大量肌原纤维、Z小体、多形性线粒体和分泌颗粒。供体的实际年龄是培养成人组织的一个重要因素,较年轻的组织成功率更高。该方法为成人心肌细胞的体外研究提供了一种手段,并为合适的心房组织使用提供了指导。