title: Efficient Generation of Glucose-Responsive Beta Cells from Isolated GP2+Human Pancreatic Progenitors.
abstract: Stem cell-based therapy for type 1 diabetes would benefit from implementation of a cell purification step at the pancreatic endoderm stage. This would increase the safety of the final cell product, allow the establishment of an intermediate-stage stem cell bank, and provide a means for upscaling β cell manufacturing. Comparative gene expression analysis revealed glycoprotein 2 (GP2) as a specific cell surface marker for isolating pancreatic endoderm cells (PECs) from differentiated hESCs and human fetal pancreas. Isolated GP2+PECs efficiently differentiated into glucose responsive insulin-producing cells in vitro. We found that in vitro PEC proliferation declines due to enhanced expression of the cyclin-dependent kinase (CDK) inhibitors CDKN1A and CDKN2A. However, we identified a time window when reducing CDKN1A or CDKN2A expression increased proliferation and yield of GP2+PECs. Altogether, our results contribute tools and concepts toward the isolation and use of PECs as a source for the safe production of hPSC-derived β cells.
authors.affiliation: The Danish Stem Cell Center (DanStem), Faculty of Health Sciences, University of Copenhagen, 2200 Copenhagen N, Denmark; Lund Stem Cell Center, Department of Laboratory Medicine, Lund University, BMC, B10, 22184 Lund, Sweden.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Ameri
authors.affiliation: Center for Genomic Medicine, Copenhagen University Hospital, Rigshospitalet, Blegdamsvej 9, 2100 Copenhagen, Denmark.
authors.forename: Rehannah
authors.initials: R
authors.lastname: Borup
authors.affiliation: The Danish Stem Cell Center (DanStem), Faculty of Health Sciences, University of Copenhagen, 2200 Copenhagen N, Denmark.
authors.forename: Christy
authors.initials: C
authors.lastname: Prawiro
authors.affiliation: INSERM U1016, University Paris-Descartes, Cochin Institute, 75014 Paris, France.
authors.forename: Cyrille
authors.initials: C
authors.lastname: Ramond
authors.affiliation: The Danish Stem Cell Center (DanStem), Faculty of Health Sciences, University of Copenhagen, 2200 Copenhagen N, Denmark.
authors.forename: Karen A
authors.initials: KA
authors.lastname: Schachter
authors.affiliation: INSERM U1016, University Paris-Descartes, Cochin Institute, 75014 Paris, France.
authors.forename: Raphael
authors.initials: R
authors.lastname: Scharfmann
authors.affiliation: The Danish Stem Cell Center (DanStem), Faculty of Health Sciences, University of Copenhagen, 2200 Copenhagen N, Denmark; Lund Stem Cell Center, Department of Laboratory Medicine, Lund University, BMC, B10, 22184 Lund, Sweden. Electronic address: semb@sund.ku.dk.
authors.forename: Henrik
authors.initials: H
authors.lastname: Semb
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050763:Cyclin-Dependent Kinase Inhibitor p18 / Q000235:genetics / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics / Q000378:metabolism; D003922:Diabetes Mellitus, Type 1 / Q000628:therapy; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D004707:Endoderm / Q000166:cytology; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D055106:Genome-Wide Association Study; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D015415:Biomarkers; C493956:CDKN1A protein, human; C000614131:CDKN2A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050763:Cyclin-Dependent Kinase Inhibitor p18; D050759:Cyclin-Dependent Kinase Inhibitor p21; C489339:GP2 protein, human; D058851:GPI-Linked Proteins; D007328:Insulin; D005947:Glucose
keywords: CDKN1A; CDKN2A; GP2; cell surface marker; cell-cycle regulators; differentiation; human embryonic stem cells; insulin-producing beta cells; pancreatic progenitors; type 1 diabetes