新生Wistar大鼠心肌细胞的分离培养与鉴定

Isolation and Culture of Cadiocytes from Neonatal Wistar Rats

本试验取新生24h内的Wistar大鼠心肌组织,无菌剪碎后采用胰酶消化法分离细胞,用含20%胎牛血清的DMEM/F12培养基培养,逐日在倒置相差显微镜下观察。结果于培养后48h即可观察到部分细胞搏动;72h后,细胞逐渐展开,搏动频率在60～70次/min,从而初步确立培养的细胞为心肌细胞;进一步采用心肌细胞特异性蛋白(cTnT)抗体和α肌动蛋白(α-actin)单克隆抗体分别对培养的心肌细胞进行免疫荧光及免疫组织化学鉴定,结果均呈阳性。

To study the method of primary culture of cadiocytes in vitro using tissue explants of cardiac muscles isolated from neonatal Wistar rats. The hearts of the neonatal rats were removed and cut into small pieces, using the method of trypsinization isolated cell and cultivated in DMEM/F12 culture medium with 20 V00 culture medium, daily observation under in- verted phase contrast microscope. The results observed that several cells had the characteristic of rhythmic contraction after cultured 48 h; after 72 h, the cells gradually spread and the rhythmic contraction was 60 to 70 times/rain and they were con- firmed to be cadiocytes. Furthermore the cadiocytes were positive, which were identified by immunofluorescence with cTnT antibody and immunohistochemical strain of a-actin monoelonal antibody. This method of tissue culture is simple by which more single eadiocytes with good functional status can be obtained.