成年小鼠肺上皮干细胞/祖细胞的克隆培养

Clonal culture of adult mouse lung epithelial stem/progenitor cells.

作者信息

McQualter Jonathan L, Bertoncello Ivan

机构信息

Lung Health Research Centre, Department of Pharmacology and Therapeutics, University of Melbourne, Grattan Street, Parkville, VIC, 3010, Australia,

出版信息

Methods Mol Biol. 2015;1235:231-41. doi: 10.1007/978-1-4939-1785-3_17.

DOI:10.1007/978-1-4939-1785-3_17

PMID:25388397

Abstract

Clonal culture of stem cells is crucial for their identification, and the characterization of the cellular and molecular mechanisms that regulate their proliferation and differentiation. In the adult mouse lung, epithelial stem/progenitor cells are defined by the phenotype CD45(neg) CD31(neg) EpCAM(pos) CD104(pos) CD24(low). Here we describe a tissue dissociation and flow cytometry strategy for the detection and isolation of adult mouse lung epithelial stem/progenitor cells, and a three-dimensional colony-forming assay for their clonal culture in vitro.

摘要

干细胞的克隆培养对于其鉴定以及调节其增殖和分化的细胞与分子机制的表征至关重要。在成年小鼠肺中，上皮干细胞/祖细胞由CD45(阴性)CD31(阴性)EpCAM(阳性)CD104(阳性)CD24(低表达)的表型定义。在此，我们描述了一种用于检测和分离成年小鼠肺上皮干细胞/祖细胞的组织解离和流式细胞术策略，以及一种用于其体外克隆培养的三维集落形成测定法。