title: Targeting pancreatic progenitor cells in human embryonic stem cell differentiation for the identification of novel cell surface markers.
abstract: New sources of beta cells are needed in order to develop cell therapies for patients with diabetes. An alternative to forced expansion of post-mitotic beta cells is the induction of differentiation of stem-cell derived progenitor cells that have a natural self-expansion capacity into insulin-producing cells. In order to learn more about these progenitor cells at different stages along the differentiation process in which they become progressively more committed to the final beta cell fate, we took the approach of identifying, isolating and characterizing stage specific progenitor cells. We generated human embryonic stem cell (HESC) clones harboring BAC GFP reporter constructs of SOX17, a definitive endoderm marker, and PDX1, a pancreatic marker, and identified subpopulations of GFP expressing cells. Using this approach, we isolated a highly enriched population of pancreatic progenitor cells from hESCs and examined their gene expression with an emphasis on the expression of stage-specific cell surface markers. We were able to identify novel molecules that are involved in the pancreatic differentiation process, as well as stage-specific cell markers that may serve to define (alone or in combination with other markers) a specific pancreatic progenitor cell. These findings may help in optimizing conditions for ultimately generating and isolating beta cells for transplantation therapy.
authors.affiliation: Stem Cell Therapeutics Ltd, Jerusalem, Israel.
authors.forename: Bettina
authors.initials: B
authors.lastname: Fishman
authors.forename: Hanna
authors.initials: H
authors.lastname: Segev
authors.forename: Oded
authors.initials: O
authors.lastname: Kopper
authors.forename: Jonathan
authors.initials: J
authors.lastname: Nissenbaum
authors.forename: Margarita
authors.initials: M
authors.lastname: Schulman
authors.forename: Nissim
authors.initials: N
authors.lastname: Benvenisty
authors.forename: Joseph
authors.initials: J
authors.lastname: Itskovitz-Eldor
authors.forename: Danny
authors.initials: D
authors.lastname: Kitsberg
mesh_terms: D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000235:genetics; D015415:Biomarkers / Q000378:metabolism; D019759:Carbon-Nitrogen Lyases; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D003922:Diabetes Mellitus, Type 1 / Q000628:therapy; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D050417:Insulin-Secreting Cells / Q000637:transplantation; D008565:Membrane Proteins / Q000378:metabolism*; D019884:Nitrogenous Group Transferases / Q000096:biosynthesis / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D010179:Pancreas / Q000166:cytology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D055760:SOXF Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D059467:Transcriptome; D019076:Transgenes
chemical_list: D029681:Arabidopsis Proteins; D015415:Biomarkers; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C109188:SOX17 protein, human; D055760:SOXF Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019884:Nitrogenous Group Transferases; D019759:Carbon-Nitrogen Lyases; C505161:At2g38230 protein, Arabidopsis