摘要
目的建立经济适用的小鼠胚胎心肌细胞的分离、培养和鉴定方法。方法取不同胚龄小鼠心脏，采用不同方法进行消化、接种、体外增生、鉴定。结果用0．125％胰蛋白酶和0．02％EDTA混合消化液，短时间、分步消化孕12d鼠胚的心脏。将获得的细胞悬液接种于胎牛血清包被的培养皿中，采用pH值为7．2～7．4的HAM’SF12加20％胎牛血清培养液进行培养，所获得的细胞经PAS反应及免疫组化鉴定显示约80％的细胞为心肌细胞。结论建立的鼠胚心肌细胞培养模型可进行胚胎心脏发育毒性的体外研究。
Objective To establish the model of primary cultured mice embryonic cardiomyocytes, which is adapted to isolation, culture and identification. Methods To isolate hearts from different day gestational age mice, then digested, seeded, proliferation in vitro and identified in different ways. Results Repeat digestion with 0. 125 % trypsin and 0.02 % EDTA , then in the form of cell suspension seeded in tissue culture plate precoated with fetal bovine serum, and added with Ham F12 containing 20 % FBS （pH 7.2 -7.4）, and finally identified cells with P AS staining and immunocytochemical methods, which proved the rate between cardiomyocytes and cultured cells was about 80%. Conclusion The model of cultured mice embryonic cardiomyocytes has highly effective.
出处
《首都医科大学学报》
CAS
2006年第5期604-606,共3页
Journal of Capital Medical University
基金
国家自然科学基金重点项目(30030120)资助项目
