链霉蛋白酶消化时长对气液相培养原代小鼠气道上皮细胞的影响  

Optimalpronase digestion time for culture and differentiation of primary mouse tracheal epithelial cells

作　　者：高璇 徐菲[2] 许先荣[1] GAO Xuan;XU Fei;XU Xianrong(Department of Respiratory Medicine,Tongde Hospital of Zhejiang Province,Hangzhou 310012,China)

机构地区：[1]浙江省立同德医院呼吸内科,杭州310012 [2]浙江大学医学院附属第一医院内科病房

出　　处：《浙江医学》2019年第13期1363-1366,I0006,共5页Zhejiang Medical Journal

摘　　要：目的观察链霉蛋白酶消化时长对气液相(ALI)培养原代小鼠气道上皮细胞(MTEC)数量、存活率及纯度的影响。方法从4组小鼠获得并培养的原代MTEC,分别进行链霉蛋白酶消化6、12、18、24h,比较4组细胞的数量、存活率及纯度。结果链霉蛋白酶消化6、12、18、24h,分别获得细胞(0.57±0.03)、(1.05±0.22)、(1.13±0.22)、(1.28±0.39)×10^5个,差异有统计学意义(P<0.05);细胞存活率分别为(97.65±2.26)%、(96.83±0.76)%、(93.46±2.32)%、(91.56±2.99)%,差异有统计学意义(P<0.05),其中6h组、12h组均明显高于18h组、24h组(均P<0.05);细胞纯度分别为(69.6±22.6)%、(90.4±5.1)%、(90.3±2.7)%、(88.7±1.9)%,差异有统计学意义(P<0.05),其中12h组、18h组均明显高于6h、24h组(均P<0.05)。结论ALI培养原代MTEC时,链霉蛋白酶消化12h可获得最优的细胞数、细胞存活率及细胞纯度。Objective To test the optimalpronase digestion time for culture and differentiation of primary mouse tracheal epithelial cells(MTECs).Methods MTECs were cultured with different pronasedigestion times(6,12,18,24 h),thecell yield,viability and the purity of differentiated cells were compared.Results The optimal digest time was 6 h,with a cell yield of(0.57±0.03)×10^5 per mouse,cell viability of(97.65±2.26)%and cell purity of(69.6±22.6)%after differentiation.The optimal digest time was 12 h,with a cell yield of(1.05±0.22)×10^5 per mouse,cell viability of(96.83±0.76)%and cell purity of(90.4±5.1)%after differentiation.The optimal digest time was 18 h,with a cell yield of(1.13±0.22)×10^5 per mouse,cell viability of(93.46±2.32)%and cell purity of(90.3±2.7)%after differentiation.The optimal digest time was 24 h,with a cell yield of(1.28±0.39)×10^5 per mouse,cell viability of(91.56±2.99)%and cell purity of(88.7±1.9)%after differentiation.The cell yield,viability and purity were significantly different among three assays.Conclusion The study suggests that 12 h may be the optimalpronase digestion time for culture and differentiation of mouse tracheal epithelial cells.

关 键 词：上皮细胞 细胞培养方法 培养基 链霉蛋白酶 消化时长 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]