Postnatal Pancreas of Mice Contains Tripotent Progenitors Capable of Giving Rise to Duct, Acinar, and Endocrine Cells In Vitro.
Postnatal pancreas is a potential source for progenitor cells to generate endocrine β-cells for treating type 1 diabetes. However, it remains unclear whether young (1-week-old) pancreas harbors multipotent progenitors capable of differentiating into duct, acinar, and endocrine cells. Laminin is an extracellular matrix (ECM) protein important for β-cells' survival and function. We established an artificial extracellular matrix (aECM) protein that contains the functional IKVAV (Ile-Lys-Val-Ala-Val) sequence derived from laminin (designated aECM-lam). Whether IKVAV is necessary for endocrine differentiation in vitro is unknown. To answer these questions, we cultured single cells from 1-week-old pancreas in semi-solid media supplemented with aECM-lam, aECM-scr (which contains a scrambled sequence instead of IKVAV), or Matrigel. We found that colonies were generated in all materials. Individual colonies were examined by microfluidic reverse transcription-polymerase chain reaction, immunostaining, and electron microscopy analyses. The majority of the colonies expressed markers for endocrine, acinar, and ductal lineages, demonstrating tri-lineage potential of individual colony-forming progenitors. Colonies grown in aECM-lam expressed higher levels of endocrine markers Insulin1, Insulin2, and Glucagon compared with those grown in aECM-scr and Matrigel, indicating that the IKVAV sequence enhances endocrine differentiation. In contrast, Matrigel was inhibitory for endocrine gene expression. Colonies grown in aECM-lam displayed the hallmarks of functional β-cells: mature insulin granules and glucose-stimulated insulin secretion. Colony-forming progenitors were enriched in the CD133(high) fraction and among 230 micro-manipulated single CD133(high) cells, four gave rise to colonies that expressed tri-lineage markers. We conclude that young postnatal pancreas contains multipotent progenitor cells and that aECM-lam promotes differentiation of β-like cells in vitro.
1 Irell & Manella Graduate School of Biological Sciences, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.|2 The Department of Translational Research & Cellular Therapeutics, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.
Nadiah
N
Ghazalli
3 Department of Bioengineering, California Institute of Technology , Pasadena, California.
Alborz
A
Mahdavi
2 The Department of Translational Research & Cellular Therapeutics, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.
Tao
T
Feng
2 The Department of Translational Research & Cellular Therapeutics, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.
Liang
L
Jin
4 Division of Chemistry and Chemical Engineering, California Institute of Technology , Pasadena, California.
Mark T
MT
Kozlowski
2 The Department of Translational Research & Cellular Therapeutics, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.
Jasper
J
Hsu
1 Irell & Manella Graduate School of Biological Sciences, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.|2 The Department of Translational Research & Cellular Therapeutics, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.
Arthur D
AD
Riggs
4 Division of Chemistry and Chemical Engineering, California Institute of Technology , Pasadena, California.
David A
DA
Tirrell
1 Irell & Manella Graduate School of Biological Sciences, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.|2 The Department of Translational Research & Cellular Therapeutics, Diabetes & Metabolism Research Institute at City of Hope , Duarte, California.
H Teresa
HT
Ku
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D003094:Collagen / Q000378:metabolism; D004338:Drug Combinations; D004703:Endocrine System / Q000166:cytology*; D007328:Insulin / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology*; D007797:Laminin / Q000378:metabolism; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology*; D010179:Pancreas / Q000166:cytology*; D011509:Proteoglycans / Q000378:metabolism
D004338:Drug Combinations; D007328:Insulin; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen