梯度离心法分离、富集、纯化新生乳鼠心肌细胞

Gradient centrifugation for isolation and purification of neonatal rat cardiomyocytes

目的:探讨体外心肌细胞的分离与培养实验方法,建立研究心脏疾病的基础实验平台。方法:取新生小鼠心脏12只,利用胰酶和胶原酶双消化6次,利用percoll密度梯度离心法离心纯化分离细胞,贴壁培养1h,分离层纤维细胞,抗体标记分离后的心肌细胞,流式细胞仪鉴定细胞纯度。结果:分离新生小鼠的心肌细胞的纯度为91.3%。结论:本方法操作简单,获得心肌细胞的分离效率高,存活时间长。

Objective： To explore an method for in vitro isolation and culturing of myocardial cells from neonatal mouse heart,trying to provide basis for further researches on heart disease.Methods： Twelve neonatal rat hearts were processed with pancreatin and collagenase for six times;then myocardial cells were purified and isolated with percoll density gradient centrifugation.Cells were then plated and incubated for 1 h,finally the purity of antibody marked cells were evaluated with flow cytometry.Results： Flow cytometry demonstrated 91.3% purity of the neonatal rat cardiomyocytes.Conclusions： The method is simple and stable,which can also increase the efficiency of myocardial cell isolation.