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Targeting Pancreatic Progenitor Cells in Human Embryonic Stem Cell Differentiation for the Identification of Novel Cell Surface Markers

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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.

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Acknowledgments

We thank Y. Mayshar for his help in chip analysis. This research was supported by grants from JDRF and ESTOOLS.

Conflict of interest

The authors indicate no potential conflict of interest.

Author information

Authors and Affiliations

  1. Stem Cell Therapeutics Ltd, Jerusalem, Israel

    Bettina Fishman & Danny Kitsberg

  2. Stem Cell Center, Faculty of Medicine, Technion, Haifa, Israel

    Hanna Segev, Margarita Schulman & Joseph Itskovitz-Eldor

  3. Stem Cell Unit, Department of Genetics, The Hebrew University, Jerusalem, Israel

    Oded Kopper, Jonathan Nissenbaum, Nissim Benvenisty & Danny Kitsberg

  4. Department of Obstetrics and Gynecology, Rambam Health Care Campus, Haifa, Israel

    Hanna Segev & Joseph Itskovitz-Eldor

Authors
  1. Bettina Fishman
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  2. Hanna Segev
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  3. Oded Kopper
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  4. Jonathan Nissenbaum
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  5. Margarita Schulman
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  6. Nissim Benvenisty
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  7. Joseph Itskovitz-Eldor
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  8. Danny Kitsberg
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Corresponding authors

Correspondence to Nissim Benvenisty or Joseph Itskovitz-Eldor.

Additional information

Bettina Fishman, Hanna Segev and Oded Kopper have contributed equally.

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Supplementary Figure 1

GFP positive cells were isolated from SOX17 reporter clones and from PDX1 reporter clones by FACS sorting. Microarray analysis was performed on the GFP+ cells using an Affymetrix GeneChip Human Gene ST1.0 array. A. Comparison between the transcription factors and secreted ligands with the highest variation between the various groups. (I)- Highly expressed in SOX17. (II)- Highly expressed in PDX1. B. Relative expression of published surface markers in undifferentiated hESCs, sorted SOX17 cells, sorted PDX1 cells and pancreas. (PDF 509 kb)

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Fishman, B., Segev, H., Kopper, O. et al. Targeting Pancreatic Progenitor Cells in Human Embryonic Stem Cell Differentiation for the Identification of Novel Cell Surface Markers. Stem Cell Rev and Rep 8, 792–802 (2012). https://doi.org/10.1007/s12015-012-9363-x

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