不同方法培养SD大鼠心肌干细胞的比较

Comparison study of different culture methods of rat cardiac stem cells in vitro

目的比较不同方法分离和体外扩增SD大鼠心肌干细胞的特点。方法分离出SD大鼠心肌组织,分别采用组织块培养法、改良组织块培养法进行心肌干细胞原代培养;分别采用差速消化法、快速消化洗脱法进行传代。结果成纤维样细胞首先从组织块爬出,其后小、圆、亮细胞迁移至成纤维样细胞层上并长成大量集落。改良组织块培养法小、圆、亮细胞较易爬出,快速消化洗脱法传代可去除大量成纤维样细胞混杂。结论改良组织块培养法是一种良好的大鼠心肌干细胞培养方法,快速消化洗脱法传代纯化率较高。

Objective To compare the efficacies of 2 different culture methods of rat cardiac stem cells （CSCs） in vitro. Methods The myocardial tissues were dissected for primary culture of CSCs by methods of explants and improved explants, respectively, and passaged by trypsin of different durations. Results Fibroblast - like cells were first crawled out from adherent explants, afterward migration of small and phase - bright cells on fibroblast - like cell layer was observed in morphologic study. More small and phase - bright cells were discovered using improved explants method. Meanwhile, 0. 05% trypsin washing for 30 seconds was proved to have the most efficient purification method. Conclusion Improved explants culture is a proven culture method for rat CSCs, with rapid purification after trypsin digestion.