A Quantitative Assay for Insulin-expressing Colony-forming ProgenitorsMichael Winkler 1, Nancy Trieu 2, Tao Feng 2, Liang Jin 2, Stephanie Walker 2, Lipi Singh 2, Hsun Teresa Ku 2,3
1Department of Biotechnology & Bioinformatics, California State University Channel Islands, 2Department of Diabetes, Endocrinology and Metabolism, Beckman Research Institute of City of Hope, 3The Irell & Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope
A three-dimensional clonogenic assay that allows pancreatic-like progenitors to differentiate into insulin-expressing colonies is described. This method takes advantage of semi-solid media containing methylcellulose, Matrigel and growth factors, in which single progenitors proliferate and differentiate in vitro, permitting quantification of the number of functional progenitors in a population.
