Bertoncello Ivan, McQualter Jonathan
Lung Regeneration Laboratory, The Department of Pharmacology, University of Melbourne, Victoria, Australia.
Curr Protoc Stem Cell Biol. 2011 Jan;Chapter 2:Unit 2G.1. doi: 10.1002/9780470151808.sc02g01s16.
Adult mouse lung epithelial stem/progenitor cells (EpiSPC) can be defined in vitro as epithelial colony-forming units that are capable of self-renewal, and which when co-cultured with lung mesenchymal stromal cells (MSC) are able to give rise to differentiated progeny comprising mature lung epithelial cells. This unit describes a protocol for the prospective isolation and in vitro propagation and differentiation of adult mouse lung EpiSPC. The strategy used for selection of EpiSPC and MSC from adult mouse lung by enzymatic digestion and flow cytometry is based on the differential expression of CD45, CD31, Sca-1, EpCAM, and CD24. The culture conditions required for the differentiation (co-culture with MSC) and expansion (stromal-free culture with FGF-10 and HGF) of EpiSPC are described.
成年小鼠肺上皮干细胞/祖细胞（EpiSPC）在体外可定义为具有自我更新能力的上皮集落形成单位，当与肺间充质基质细胞（MSC）共培养时，能够产生包含成熟肺上皮细胞的分化后代。本单元描述了一种用于成年小鼠肺EpiSPC前瞻性分离、体外增殖和分化的方案。通过酶消化和流式细胞术从成年小鼠肺中选择EpiSPC和MSC的策略基于CD45、CD31、Sca-1、EpCAM和CD24的差异表达。描述了EpiSPC分化（与MSC共培养）和扩增（用FGF-10和HGF进行无基质培养）所需的培养条件。
