The long term goal of the studies in this proposal is to understand the biology of pancreatic ? cell like progenitors derived from embryonic stem (ES) cells, with the intent of potentially using these cells for clinical applications such as islet transplantation for type I diabetics. Of particular relevance to this application, we have developed a novel colony assay for ? like progenitors in vitro. This colony assay or culture system allows dissociated single cells to develop into insulin-expressing colonies after 8 days of culture in a semi-solid media. Such assay measures the function, rather than phenotype, of ? like progenitors. In addition, this assay can be used to quantitate the number of ? like progenitors or the insulin-expressing colony forming units (ICFUs) in a given population. With this tool available, we propose to study fundamental properties of ICFUs derived from murine ES cells. We hypothesize that ICFUs can be enriched by cell surface markers, are expandable in number, and can be further matured in vitro. The studies outlines in the proposal will 1) sort and enrich ICFUs using cell surface markers, 2) expand ICFUs using exogenous growth factors, and 3) derive mature insulin-expressing colonies from ICFUs. These studies may lead to the development of novel methods for producing large numbers of functional insulin-secreting cells for potential therapeutic applications, and the possible identification of multipotential pancreatic endocrine progenitors or stem cells in vitro.

Public Health Relevance

A novel in vitro clonogenic assay that measures the activities of pancreatic beta cell like progenitors will be used to study the biological properties of these cells. We will test whether the murine embryonic stem cell derived beta cell like progenitors can be enriched by cell surface markers, expanded by exogenous factors and matured into functional glucose-sensing insulin-secreting cells. These studies may lead to the development of efficient methods for cell replacement therapy for Type I diabetics, and the possible identification of multipotential pancreatic endocrine progenitors or stem cells in vitro.

National Institute of Health (NIH)
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Research Project (R01)
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Cellular Aspects of Diabetes and Obesity Study Section (CADO)
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Sato, Sheryl M
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City of Hope/Beckman Research Institute
United States
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Jin, Liang; Gao, Dan; Feng, Tao et al. (2016) Cells with surface expression of CD133highCD71low are enriched for tripotent colony-forming progenitor cells in the adult murine pancreas. Stem Cell Res 16:40-53
Tremblay, Jacob R; LeBon, Jeanne M; Luo, Angela et al. (2016) In Vitro Colony Assays for Characterizing Tri-potent Progenitor Cells Isolated from the Adult Murine Pancreas. J Vis Exp :
Ghazalli, Nadiah; Mahdavi, Alborz; Feng, Tao et al. (2015) Postnatal Pancreas of Mice Contains Tripotent Progenitors Capable of Giving Rise to Duct, Acinar, and Endocrine Cells In Vitro. Stem Cells Dev 24:1995-2008
Jin, Liang; Feng, Tao; Chai, Jing et al. (2014) Colony-forming progenitor cells in the postnatal mouse liver and pancreas give rise to morphologically distinct insulin-expressing colonies in 3D cultures. Rev Diabet Stud 11:35-50
Jin, Liang; Feng, Tao; Zerda, Ricardo et al. (2014) In vitro multilineage differentiation and self-renewal of single pancreatic colony-forming cells from adult C57BL/6 mice. Stem Cells Dev 23:899-909
Fu, Xianghui; Jin, Liang; Wang, Xichun et al. (2013) MicroRNA-26a targets ten eleven translocation enzymes and is regulated during pancreatic cell differentiation. Proc Natl Acad Sci U S A 110:17892-7
Jin, Liang; Feng, Tao; Shih, Hung Ping et al. (2013) Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel. Proc Natl Acad Sci U S A 110:3907-12
Winkler, Michael; Trieu, Nancy; Feng, Tao et al. (2011) A quantitative assay for insulin-expressing colony-forming progenitors. J Vis Exp :e3148
Chen, Chialin; Chai, Jing; Singh, Lipi et al. (2011) Characterization of an in vitro differentiation assay for pancreatic-like cell development from murine embryonic stem cells: detailed gene expression analysis. Assay Drug Dev Technol 9:403-19
Kuroda, Akio; Rauch, Tibor A; Todorov, Ivan et al. (2009) Insulin gene expression is regulated by DNA methylation. PLoS One 4:e6953