中国动脉硬化杂志2011，Vol.19Issue(4)：361-366,6.
一套系统培养鼠主动脉血管壁细胞简单可靠的方法
The Simple and Reliable Methods of Systematically Culturing Rat Aortic Wall Cells
摘要
Abstract
Aim To discuss a set of system to culture vascular smooth muscle cell (VSMC), vascular adventitial fibroblast (VAF) and vascular endothelial cell (VEC) from murine aorta, which are simple, reliable and easy to replicate. Methods The primary culture of VSMC, VAF and VEC used the tissue adherent method, trypsin digestion for passage transfer. The cells were purified by differential adherent and natural growth purification. Myofibroblast (MF) was transformed from VAF by growth factor-βl (TGF-β1) induced. Phase contrast microscope and immunocytochemistry staining were used to identify the morphological and the immunological characteristics, respectively. Results The tissues and cells activity were excellent. The growth cycle of primary VSMC/VAF and VEC were 10 ～ 12 days and 12 ～ 14 days, respectively. And passage cycle was 7 ～ 10 days. After being purified and subculture, the cells purity achieved 95％ ～ 100％. The growth characteristics of VSMC assumed the model "peak and valley", and with anti a-SMA ( + ) / Vimentin ( - ); the VEC assumed "pebble-like" appearance, and anti CD31 ( + )/a-SMA ( - ); the staining of VAF were Vimentin ( + )/a-SMA ( - ), while the induction MF were Vimentin ( + )/a-SMA ( + ). The primary cells were successfully passaged more than 10 generations, without growth vigor decreasing. Conclusion We have established a set of system methods to culture the primary VEC, VSMC, VAF and MF of rat aorta origin, the advantages of our approach are simple, reliable and easy to replicate, while the cells have high purity and excellent biological activity. This method is still suitable for culturing the ingredient cell of mouse aorta vessel wall.关键词
主动脉/原代细胞培养/血管平滑肌细胞/外膜成纤维细胞/肌成纤维细胞Key words
Aorta/ Primary Cell Culture/ Vascular Smooth Muscle Cell/ Adventitial Fibroblast/ Myofibroblast分类
生物科学引用本文复制引用
宋方,吴强,陆德琴,袁军,杨永耀,谭洪文..一套系统培养鼠主动脉血管壁细胞简单可靠的方法[J].中国动脉硬化杂志,2011,19(4):361-366,6.基金项目
国家自然科学基金资助项目(30871003) （30871003）
贵州省优秀科技教育人才省长基金项目(黔省专合字[2009]30号) （黔省专合字[2009]30号）
