摘要
目的比较人脐动脉平滑肌细胞(HUASMC)两种原代培养方法的优劣.方法分离脐动脉,剥离中膜,将其剪成小块,贴块法将小块直接接种于培养瓶中;酶解法①单用胶原酶Ⅰ(0.2%)分别消化5小时、8小时、24小时后接种于培养瓶,②分别用胶原酶Ⅰ(0.2%)消化3小时和胰蛋白酶(0.25%)消化2小时,接种于培养瓶.结果贴块法6天可以见到细胞从组织块周围游出,20～30天后可以进行传代;单用胶原酶Ⅰ消化24小时可以见到少量细胞,但由于细胞数少,无法进行传代;单用胶原酶Ⅰ消化5小时和8小时以及用胶原酶Ⅰ和胰蛋白酶联合消化均未见到贴壁生长的细胞.结论HUASMC原代培养贴块法较酶解法简便经济且成功率较高.
Objective： To compare two methods of human umbilical artery smooth muscle cell （HUASMC） primary culturing. Methods： Human umbical artery was isolated, the middle layer was separated and cut into small pieces and then cultivated in DMEM/F12 culture medium with 20% fetal calf serum （FCS） by explant method; by enzymatically dispersed method, the small pieces were treated with collagenase Ⅰ（0.2%） for 5, 8, 24 hours, or first treated with collagenase Ⅰ（0.2%） for 2 hours and with trypsin （0.25%） for 3 hours, then cultivated in DMEM/F12 culture medium with 20% FCS. Results： Cells could be seen around the small pieces after 6 days by explant method, and be subeultured after 20～30 days; while only a few cells could be seen by enzymatically dispersed method with collagenase Ⅰ for 24 hours, but couldn＇t be subcultured because of the small amount of cells; no cells could be seen by other methods. Conclusion： Explant method is much easier more economical in cultivating HUASMC.
出处
《泰山医学院学报》
CAS
2005年第3期187-189,共3页
Journal of Taishan Medical College
基金
山东省自然科学基金(NO.Z2002C40)
关键词
人脐动脉平滑肌细胞
原代培养
贴块法
酶解法
human umbilical artery smooth muscle cell
primary culture
explant method
enzymatically dispersed method
