新生大鼠心肌细胞和心肌成纤维细胞的同时分离  

Simultaneous isolation of myocardial cells and cardiac fibroblasts from neonatal rats

作　　者：施雨露[1] 李晓辕[2] 曹美娜[3] 于姝媛[3] 王苹[3] 

机构地区：[1]吉林大学白求恩医学院病原免疫细胞遗传实验中心,吉林省长春市130021 [2]吉林大学第一医院呼吸科,吉林省长春市130021 [3]吉林大学第一医院耳鼻咽喉-头颈外科,吉林省长春市130021

出　　处：《中国组织工程研究》2013年第24期4414-4420,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:利用心肌组织中不同细胞的黏附特性差异,通过优化心肌细胞的培养条件,一次分离同时获得心肌和心肌成纤维细胞。目的:探讨新生大鼠心肌和心肌成纤维细胞同时分离的有效方法。方法:采用低浓度胰蛋白酶冷消化结合胶原酶复合消化,差速分离心肌和心肌成纤维细胞。通过使用不同种类的血清改良心肌细胞培养条件,免疫荧光鉴定心肌细胞和心肌成纤维细胞纯度。结果与结论:心肌培养48h时TroponinT阳性细胞率为89.3%。第2代的心肌成纤维细胞培养波形蛋白阳性细胞率为93.6%。含马血清的培养基组心肌细胞阳性率明显高于牛血清培养组(P<0.05)。实验应用的新生大鼠心肌细胞分离培养方法,心肌细胞和心肌成纤维细胞纯度高、活性好,且能同时获得心肌细胞和心肌成纤维细胞。BACKGROUND： Myocardial cells and cardiac fibroblasts are simultaneously isolated through optimizing the culture conditions of myocardial cells, by use of the variations of adhesion properties of myocardial cells. OBJECTIVE： To explore the co-separation method for myocardial cells and cardiac fibroblasts in newborn ratsMETHODS： Myocardial tissue in newborn rats was digested with low concentration trypsin at cold conditions and with collagenase II at 37 ℃. The myocardial cells and cardiac fibroblasts were isolated with differential centrifugation method, and myocardial cell culture conditions were improved using different kinds of serum. Immunofluorescence staining was used to evaluate the purity of myocardial cells and cardiac fibroblasts. RESULTS AND CONCLUSION： The myocardial Troponin T positive cell rate was 89.3% at culture 48 hours, and myocardial fibroblast vimentin positive cell. rate was 93.6% at passage 2. The percentage of positive myocardial cells cultured with medium containing horse serum was obviously higher than that with bovine serum culture （P 〈 0.05）. This isolation method of rat myocardial cells can gain myocardial cells and cardiac fibroblasts at the same time, with high pudty and good activity.

关 键 词：组织构建 心脏组织构建 心肌细胞 心肌成纤维细胞 胰蛋白酶 胶原酶 复合消化 细胞培养 马 血清 牛血清 增殖 分化 

分 类 号：R318[医药卫生—生物医学工程]