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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
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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.
作者 高璇 徐菲 许先荣 GAO Xuan;XU Fei;XU Xianrong(Department of Respiratory Medicine,Tongde Hospital of Zhejiang Province,Hangzhou 310012,China)
出处 《浙江医学》 CAS 2019年第13期1363-1366,I0006,共5页 Zhejiang Medical Journal
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 Epithelial cell Cell culture techniques Culture media Pronase Digestion time
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  • 1张景熙,李强,白冲,刘忠令,黄盛东,龚德军.经纤维支气管镜刷检人支气管上皮细胞的原代培养[J].第二军医大学学报,2004,25(7):791-792. 被引量:7
  • 2Shao-hengHE JianZHENG Ming-keDUAN.Induction of mucin secretion from human bronchial tissue and epithelial cells by rhinovirus and lipopolysaccharide[J].Acta Pharmacologica Sinica,2004,25(9):1176-1181. 被引量:4
  • 3Yaghi A, Zaman A, Dolovich M. Primary human bronchial epithelial ceils grown from explants [J]. J Vis Exp, 2010,26 (37). pii: 1789. doi: 10.3791/1789.
  • 4James L. Lordan, Fabio Bucchieri, 2 Audrey Richter, et al. Cooperative Effects of Th2 Cytokines and Allergen on Normal and Asthmatic Bronchial Epithelial Cells [J]. Immunol, 2002, 169 : 407-414.
  • 5秦晓群,向阳,刘持,谭宇蓉,屈飞,彭丽花,朱晓琳,秦岭.支气管上皮细胞在气道高反应中的作用(英文)[J].生理学报,2007,59(4):454-464. 被引量:13
  • 6司徒镇强 吴军正.细胞培养[M].西安:世界图书出版社,2003.66-67.
  • 7Stenn KS, Link R, Moellmann G, et al. Dispase, a neutral protease from bacillus polymyxa, is a powerful fibronectinase and type Ⅳ collagenase.J Invest Dermatol, 1989, 93:287-290.
  • 8Baeza-Squiban A,Delcher L,Kuketi R.Responses of the Rabbit Tracheal Epithelium In Vitro to H2O2-induced Oxidative Stress[J].Toxicol Vitro,2000,14(2):159-167.
  • 9Brian WZ,Joachim A,Rainer S,et al.The characterisation of human respiratory epithelial cells cultured on resorbable scaffolds:first steps towards a tissue engineered tracheal replacement [J].Biomaterials,2002,23(6):1425-1438.
  • 10Theresa FY,Eileen LT,Barbara ZE,et al.A tissue culture system to study respiratory ciliary epithelial adherence of selected swine mycoplasmas[J].Veterinary Microbiol,2000,71(3-4):269-279.

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