【摘要】 为探讨雏鸡心肌细胞的分离、培养方法,采用胶原酶重复多次消化心肌组织,收集每次消化的上清液用含20%胎牛血清的DMEM/F12培养基终止消化,离心沉淀细胞后再加入培养基混悬细胞。采用差速贴壁法和化学法纯化心肌细胞后置CO2培养箱孵育,并对培养的心肌细胞进行免疫组化鉴定。结果显示,未贴壁时心肌细胞呈圆形,培养12～24h心肌细胞开始贴壁生长,细胞伸出伪足呈梭形、多角形,3～4d后形成细胞簇,5～7d后细胞汇合成片。免疫组化结果表明α-SA抗原表达阳性,证明培养的细胞为心肌细胞,应用酶消化法可简便、快速地获得大量活力高的雏鸡心肌细胞。更多还原

【Abstract】 To find an easy way to isolate and culture myocardial cells of chickens,the hearts were digested by collagenase typeⅠseveral times and then the supernatant was collected. The digestion was stopped by adding DMEM/F12 supplemented with 20% FBS. Cardiocytes was precipitated by centrifugation,and then added the medium to cardiocytes. Cells were preplated to reduce fibroblast content,and were treated with 5-bromodeoxyuridine (5-Brdu) for 2 days. The identification of cardiocytes was conducted by immunocytochemistry. The results showed that the cells were round before adhesion;12 to 24 hours later,the cells of fusiform and polygon began to adhere and stretch out the parapodium;3 to 4 days later,cell clusters formed;5 to 7 days later,the cells grew together flakily. The identified cells were cardiocytes. Large cardiocytes with well morphology and high livability can be convenient and quickly obtained by digesting of collagenase typeⅠ.更多还原