Tao Jing, Ma Yitong, Li Xiaomei, Chen Bangdang, Yang Yining, Ma Xiang, Liu Fen, Chen Xiaocui, Gai Mintao
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China.
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Zhonghua Xin Xue Guan Bing Za Zhi. 2014 Jan;42(1):53-6.
The present study describes an improved in vitro culture method for obtaining high purity, vital and fully functional cardiomyocytes from neonatal rat.
After cutting ventricular tissue with improved method, ventricular tissues were digested with low concentrations of trypsin overnight at 4 °C, and then underwent collagenase II digestion. Thereafter, cardiomyocytes were purified by combined differential adhesion and chemical inhibition methods.
Adherent cardiomyocytes were seen at 12 h after culture, spontaneously beating cardiomyocytes were observed at 24 h after culture, crosslinked cardiomyocytes were found at 48 h after culture, adhesion clustered cardiomyocytes were seen at 72 h after culture, dense network formed from inter-connected was evidenced together with radial arranged cell clusters and cell pseudopodia 96 h the mutual contact woven into and formed radically ordering cell clusters and island-like beating cardiomyocytes at 96 h after culture. The cell survival rate and purity were more than 98%.
Fully functional spontaneous beating cardiomyocytes can be obtained by the use of this improved primary neonatal rat cardiomyocytes culture method.
本研究描述了一种改进的体外培养方法，用于从新生大鼠中获得高纯度、有活力且功能完全的心肌细胞。
采用改进方法切割心室组织后，将心室组织在4℃用低浓度胰蛋白酶消化过夜，然后进行Ⅱ型胶原酶消化。此后，通过联合差速贴壁和化学抑制方法纯化心肌细胞。
培养12小时后可见贴壁心肌细胞，培养24小时后观察到自发搏动的心肌细胞，培养48小时后发现交联的心肌细胞，培养72小时后可见黏附聚集的心肌细胞，培养96小时后可见由相互连接形成的致密网络以及呈放射状排列的细胞簇和细胞伪足，相互接触交织形成呈放射状排列的细胞簇和岛状搏动心肌细胞。细胞存活率和纯度均超过98%。
使用这种改进的新生大鼠原代心肌细胞培养方法可获得功能完全的自发搏动心肌细胞。
