title: In vitro multilineage differentiation and self-renewal of single pancreatic colony-forming cells from adult C57BL/6 mice.
abstract: In a previous study we established colony assays suitable for studying murine adult (2-4 months) pancreatic progenitor cells plated in semisolid media containing methylcellulose and extracellular matrix proteins. Using these assays, we found robust in vitro progenitor cell activities (multilineage differentiation and self-renewal) from pancreatic cells of adult mice in the CD-1 outbred background. However, it was not clear whether progenitor cell activities can be detected from inbred mice, a preferred mouse model for various genetic studies. It was also not clear whether a single cell is sufficient to self-renew. Here, we show that fluorescent activated cell sorting pancreatic CD133(+) but not CD133(-) cells from adult C57Bl/6 inbred mice are enriched for progenitor cells that self-renew and give rise to multilineage colonies in vitro. The number of cells in a colony is in proportion to its diameter. Around 60% of single handpicked 3-week-old colonies express trilineage markers, indicating most progenitors are tripotent for ductal, acinar, and endocrine lineage differentiation. Approximately 80% of primary (freshly sorted) colony-forming progenitor cells are capable of giving rise to secondary progenitors in vitro, indicating that a majority of the primary progenitors self-renew. A single cell is sufficient for self-renewal and a Wnt agonist, R-Spondin1, enhances the number of secondary progenitors from the primary progenitors. Together, our pancreatic colony assays allow quantitative analyses of progenitors at a single-cell level from inbred mice. These assays will be useful for elucidating in vitro mechanisms of pancreatic progenitor cell biology.
authors.affiliation: 1 Department of Diabetes and Metabolic Diseases Research, Beckman Research Institute of City of Hope , Duarte, California.
authors.forename: Liang
authors.initials: L
authors.lastname: Jin
authors.forename: Tao
authors.initials: T
authors.lastname: Feng
authors.forename: Ricardo
authors.initials: R
authors.lastname: Zerda
authors.forename: Ching-Cheng
authors.initials: CC
authors.lastname: Chen
authors.forename: Arthur D
authors.initials: AD
authors.lastname: Riggs
authors.forename: Hsun Teresa
authors.initials: HT
authors.lastname: Ku
mesh_terms: D000071916:AC133 Antigen; D061354:Acinar Cells / Q000502:physiology; D053687:Adult Stem Cells / Q000502:physiology*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D006023:Glycoproteins / Q000378:metabolism; D008810:Mice, Inbred C57BL; D010179:Pancreas / Q000166:cytology*; D010455:Peptides / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D015415:Biomarkers; D006023:Glycoproteins; D010455:Peptides; C000606024:Prom1 protein, mouse