Pdx1 and controlled culture conditions induced differentiation of human amniotic fluid-derived stem cells to insulin-producing clusters

So Young Chun, David L. Mack, Emily Moorefield, Se Heang Oh, Tae Gyun Kwon, Mark J. Pettenati, James J. Yoo, Paolo De Coppi, Anthony Atala, Shay Soker

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

This study investigated the differentiation of human amniotic fluid-derived stem cells (hAFSCs) into insulin-producing clusters in vitro. Adenovirally-delivered mouse Pdx1 (Ad-Pdx1) induced human Pdx1 expression in hAFSCs and enhanced the coordinated expression of downstream β-cell markers. When Ad-Pdx1-transduced hAFSCs were sequentially treated with activin A, bFGF and nicotinamide and the culture plate surface coated with poly-l-ornithine, the expression of islet-associated human mRNAs for Pdx1, Pax6, Ngn3 and insulin was increased. C-peptide ELISA confirmed that Ad-Pdx1-transduced hAFSCs processed and secreted insulin in a manner consistent with that pathway in pancreatic β-cells. To sustain the β-cell-like phenotype and investigate the effect of three-dimensional (3D) conformation on the differentiation of hAFSCs, Pdx1-transduced cells were encapsulated in alginate and cultured long-term under serum-free conditions. Over 2weeks, partially differentiated hAFSC clusters increased in size and increased insulin secretion. Taken together, these data demonstrate that ectopic Pdx1 expression initiates pancreatic differentiation in hAFSCs and that a β-cell-like phenotype can be augmented by culture conditions that mimic the stromal components and 3D geometry associated with pancreatic islets.

Original languageEnglish (US)
Pages (from-to)540-549
Number of pages10
JournalJournal of Tissue Engineering and Regenerative Medicine
Volume9
Issue number5
DOIs
StatePublished - May 1 2015
Externally publishedYes

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Keywords

  • Amniotic fluid-derived stem cells
  • Cell therapy
  • Diabetes
  • Differentiation
  • Extracellular matrix components
  • Growth factors
  • Pdx1

ASJC Scopus subject areas

  • Biomedical Engineering
  • Medicine (miscellaneous)
  • Biomaterials

Cite this

Chun, S. Y., Mack, D. L., Moorefield, E., Oh, S. H., Kwon, T. G., Pettenati, M. J., Yoo, J. J., Coppi, P. D., Atala, A., & Soker, S. (2015). Pdx1 and controlled culture conditions induced differentiation of human amniotic fluid-derived stem cells to insulin-producing clusters. Journal of Tissue Engineering and Regenerative Medicine, 9(5), 540-549. https://doi.org/10.1002/term.1631