分离培养适用于染色体分选的新生小鼠皮肤成纤维细胞  

Isolation and Culturing of Fibroblasts from Newborn Mouse Skin for Chromosome Sorting

作　　者：晁天柱[1] 徐福意 徐伟[1] 李凯[1] 周宇荀[1] 肖君华[1] 

机构地区：[1]东华大学生物科学与技术研究所,上海201620

出　　处：《东华大学学报（自然科学版）》2016年第3期390-394,共5页Journal of Donghua University(Natural Science)

基　　金：国家自然科学基金面上资助项目(31371257);上海市科委关键资助项目(12140900404)

摘　　要：采用酶消化法分离培养新生小鼠皮肤成纤维细胞(mouse skin fibroblasts,MSFs).通过噻唑蓝(MTT)比色法和流式细胞术(flow cytometry,FCM),深入探讨不同培养基和血清浓度对MSFs增殖率和同步化效率的影响.结果表明:酶消化法能快速获取大量健康、高活力的MSFs;在杜氏培养液(DMEM)传代培养条件下,P2代MSFs具有高增殖率和有丝分裂指数;地美可辛工作浓度为0.05μg/mL时,阻断同步化获取M期MSFs的效率最佳.研究结果为流式细胞术分选染色体提供实验材料和研究基础.The newborn mouse skin fibroblasts （MSFs） were isolated from the skin of newborn mouse using collagenase and trypsin digestion method. And the proliferation and synchronization rate of MSFs were measured by methylthiazolyl-tetrazolium （MTT） assay and flow cytometry （FCM） under different blood serum concentration and medium, respectively. Results show that the healthy and highly active MSFs were isolated using enzyme digestion method. In different blood serum concentration and medium, MSFs have the optimum synchronization rate with 0.05 μg/mL demecocline, and highest proliferation rate and mitoticindex under the culture of Dulbecco＇s modified eagle medium （DMEM） in passage two. And the research result provides the experimental materials and research foundations for the flow chromosome sorting.

关 键 词：成纤维细胞 噻唑蓝(MTT) 细胞同步化 流式细胞术 

分 类 号：Q952[生物学—动物学]