遵义医科大学学报2023，Vol.46Issue(12)：1153-1160,1170,9.
小鼠原代气道上皮基底细胞分离、增殖与气液界面培养诱导的定向分化
Isolation and proliferation of primary airway epithelial basal cells in mice,and directed differentiation induced by ALI culture
摘要
Abstract
Objective To explore the methods of isolation,proliferation and air-liquid interface(ALI)culture fordirected differentiation of primary mouse airway epithelial basal cells.Methods Mouse trachea were obtained aseptically,and basal cells were isolated by modified enzyme digestionand identified by immunofluorescence.The cells were passaged and observed by inverted microscope for proliferation and growth,and the expression level of KRT5 was detected by immunofluorescence.The cells were cultured at ALI,and HE staining and scan-ning electron microscopy were used to observe the directed differentiation.Results Modified enzyme digestion method and proliferation medium culture could isolate basal cells with positive KRT5 tested by immunofluores-cence.The basal cells had been passed to the eighth generation(P8),and after the fifth generation(P5),the aging of the cells increased,and the immunofluorescence intensity became weaker.In ALI culture,the HE stai-ning results showed that the P1～P3 cells have multiple high layers,and the cilia could be seen on the free sur-face of P1～P4;Scanning electron microscope results indicated that the free surface of P1～P4 cells had many specialised structures,while those structures were not so many on the free surface of P5～P8 cells.Conclusion The methods above can isolate the basal cells with high purity and activity,and the proliferated cells before P4 has good proliferation and directional differentiation ability.This work provides the cellular support for the further study of the experiments related to airway remodelling in vitro.关键词
气道基底细胞/气液界面培养/定向分化/纤毛细胞Key words
airway basal cells/air-liquid interface culture/directional differentiation/ciliated cells分类
医药卫生引用本文复制引用
王彩妹,周希,杨露,王铭轲,魏强松,顾延会,何志全..小鼠原代气道上皮基底细胞分离、增殖与气液界面培养诱导的定向分化[J].遵义医科大学学报,2023,46(12):1153-1160,1170,9.基金项目
遵义市科技计划项目[NO:遵市科合HZ字(2022)255] （2022）
遵义医科大学大学生创新创业训练计划项目(NO:ZY-DC202202235). （NO:ZY-DC202202235）
