Expansion of Human Airway Basal Stem Cells and Their Differentiation as 3D Tracheospheres [chapter]

Robert E. Hynds, Colin R. Butler, Sam M. Janes, Adam Giangreco
2016 Msphere  
Although basal cells are known to function as human airway epithelial stem cells, analysis of these cells has been limited by in vitro culture techniques that only permit minimal cell growth and differentiation. Here, we report a protocol that dramatically increases the longterm expansion of primary human airway basal cells while maintaining their genomic stability using 3T3-J2 fibroblast co-culture and ROCK inhibition using Y-27632. We also describe techniques for the differentiation and
more » ... fluorescence imaging of these expanded airway stem cells as three-dimensional tracheospheres containing basal, ciliated and mucosecretory cells. This 3D procedure allows investigation of the proximal airway epithelium under more physiologically relevant conditions than those found in traditional monolayer cultures. Together these methods represent a novel platform for improved airway stem cell growth and differentiation that is compatible with high throughput, high content translational lung research as well as human airway tissue engineering and clinical cellular therapy.
doi:10.1007/7651_2016_5 pmid:27539459 fatcat:4itwedvo3vbxfcrwgm5efovr3i