作者
Fengchao Wang, David Scoville, Xi C He, Maxime M Mahe, Andrew Box, John M Perry, Nicholas R Smith, Nan Ye Lei, Paige S Davies, Megan K Fuller, Jeffrey S Haug, Melainia McClain, Adam D Gracz, Sheng Ding, Matthias Stelzner, James CY Dunn, Scott T Magness, Melissa H Wong, Martin G Martin, Michael Helmrath, Linheng Li
发表日期
2013/8/1
期刊
Gastroenterology
卷号
145
期号
2
页码范围
383-395. e21
出版商
WB Saunders
简介
Background & Aims
Identification of intestinal stem cells (ISCs) has relied heavily on the use of transgenic reporters in mice, but this approach is limited by mosaic expression patterns and difficult to directly apply to human tissues. We sought to identify reliable surface markers of ISCs and establish a robust functional assay to characterize ISCs from mouse and human tissues.
Methods
We used immunohistochemistry, real-time reverse-transcription polymerase chain reaction, and fluorescence-activated cell sorting (FACS) to analyze intestinal epithelial cells isolated from mouse and human intestinal tissues. We compared different combinations of surface markers among ISCs isolated based on expression of Lgr5–green fluorescent protein. We developed a culture protocol to facilitate the identification of functional ISCs from mice and then tested the assay with human intestinal crypts and putative ISCs.
Results
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F Wang, D Scoville, XC He, MM Mahe, A Box, JM Perry… - Gastroenterology, 2013