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。
To find an easy way to isolate and culture myocardial cells of chickens,the hearts were digested by collagenase type I several 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 ceils were round before adhesion;12 to 24 hours later,the ceils 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 type I .
出处
《中国家禽》
北大核心
2008年第12期17-19,24,共4页
China Poultry
基金
国家自然科学基金重点资助项目(30530570)
关键词
心肌细胞
原代培养
雏鸡
cardiocyte
primary culture
chicken
