摘要
目的探讨改良联合酶消化法是否能稳定提取成年小鼠原代心脏成纤维细胞。方法 6只C57成年小鼠拉颈处死后取心脏,然后采用改良联合酶消化法即用胰酶、Ⅱ型胶原酶于37℃恒温箱内消化心脏组织6 min,加血清终止消化,然后将含有细胞的消化液冰上储存,离心提取成年小鼠原代心脏成纤维细胞,传代后采用苏木素-伊红染色及免疫荧光法检测波形蛋白、盘状结构域受体2进行形态学和免疫学鉴定。结果采用改良联合酶消化法可以稳定地提取贴壁细胞。苏木素-伊红染色可见细胞呈梭形,贴壁生长;经免疫荧光鉴定,所提取的细胞波形蛋白、盘状结构域受体2表达阳性。结论改良联合酶消化法可以稳定地提取成年小鼠原代心脏成纤维细胞。
Objective To investigate whether the modified combined enzymatic digestion method can extract primary cardiac fibroblasts from adult mice stably. Methods Six C57 adult mice were put to death by neck,and the hearts were extracted. Then,the modified combined enzymatic digestion method was adopted,in which trypsin and type Ⅱ collagenase were used to digest the heart tissues for 6 min in a constant temperature box at 37 ℃,and the digestion was terminated by adding serum. Then the digestive fluid containing cells was stored in ice,and the primary cardiac fibroblasts of adult mice were centrifugated. HE staining and immunofluorescence detection of vimentin and disc domain receptor 2( DDR2) were performed after cell passage for morphological and immunological identification. Results Adherent cells could be extracted stably by combined enzyme digestion. HE staining showed that the cells were fusiform and grew adherently to the wall. After immunofluorescence identification,the extracted cells showed positive expression of vimentin and DDR2. Conclusion The modified combined enzymatic digestion method can extract adult primary cardiac fibroblasts stably.
作者
陈洪群
郑梅
晏乘曦
黄鹏
赵兴山
CHEN Hongqun;ZHENG Mei;YAN Chengxi;HUANG Peng;ZHAO Xingshan(Civil Aviation General Hospitaly Beijing 100123, China)
出处
《山东医药》
CAS
2019年第21期38-41,共4页
Shandong Medical Journal
关键词
组织细胞提取法
改良联合酶消化法
心脏成纤维细胞
原代培养
成年小鼠
Tissue cell extraction method
combined enzymatic digestion method
cardiac fibroblasts
primary culture
adult mice
