Generation of islet-like cells from mouse gall bladder by direct ex vivo reprogramming

Raymond D. Hickey, Feorillo Galivo, Jonathan Schug, Michael A. Brehm, Annelise Haft, Yuhan Wang, Eric Benedetti, Guoqiang Gu, Mark A. Magnuson, Leonard D. Shultz, Eric Lagasse, Dale L. Greiner, Klaus H. Kaestner, Markus Grompe

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Cell replacement is an emerging therapy for type 1 diabetes. Pluripotent stem cells have received a lot of attention as a potential source of transplantable β-cells, but their ability to form teratomas poses significant risks. Here, we evaluated the potential of primary mouse gall bladder epithelial cells (GBCs) as targets for ex vivo genetic reprogramming to the β-cell fate. Conditions for robust expansion and genetic transduction of primary GBCs by adenoviral vectors were developed. Using a GFP reporter for insulin, conditions for reprogramming were then optimized. Global expression analysis by RNA-sequencing was used to quantitatively compare reprogrammed GBCs (rGBCs) to true β-cells, revealing both similarities and differences. Adenoviral-mediated expression of NEUROG3, Pdx1, and MafA in GBCs resulted in robust induction of pancreatic endocrine genes, including Ins1, Ins2, Neurod1, Nkx2-2 and Isl1. Furthermore, expression of GBC-specific genes was repressed, including Sox17 and Hes1. Reprogramming was also enhanced by addition of retinoic acid and inhibition of Notch signaling. Importantly, rGBCs were able to engraft long term in vivo and remained insulin-positive for 15. weeks. We conclude that GBCs are a viable source for autologous cell replacement in diabetes, but that complete reprogramming will require further manipulations.

Original languageEnglish (US)
Pages (from-to)503-515
Number of pages13
JournalStem Cell Research
Volume11
Issue number1
DOIs
StatePublished - Jul 2013

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Islets of Langerhans
Urinary Bladder
Epithelial Cells
Genetic Transduction
Insulin
RNA Sequence Analysis
Pluripotent Stem Cells
Teratoma
Tretinoin
Type 1 Diabetes Mellitus
Genes

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Medicine(all)

Cite this

Generation of islet-like cells from mouse gall bladder by direct ex vivo reprogramming. / Hickey, Raymond D.; Galivo, Feorillo; Schug, Jonathan; Brehm, Michael A.; Haft, Annelise; Wang, Yuhan; Benedetti, Eric; Gu, Guoqiang; Magnuson, Mark A.; Shultz, Leonard D.; Lagasse, Eric; Greiner, Dale L.; Kaestner, Klaus H.; Grompe, Markus.

In: Stem Cell Research, Vol. 11, No. 1, 07.2013, p. 503-515.

Research output: Contribution to journalArticle

Hickey, RD, Galivo, F, Schug, J, Brehm, MA, Haft, A, Wang, Y, Benedetti, E, Gu, G, Magnuson, MA, Shultz, LD, Lagasse, E, Greiner, DL, Kaestner, KH & Grompe, M 2013, 'Generation of islet-like cells from mouse gall bladder by direct ex vivo reprogramming', Stem Cell Research, vol. 11, no. 1, pp. 503-515. https://doi.org/10.1016/j.scr.2013.02.005
Hickey, Raymond D. ; Galivo, Feorillo ; Schug, Jonathan ; Brehm, Michael A. ; Haft, Annelise ; Wang, Yuhan ; Benedetti, Eric ; Gu, Guoqiang ; Magnuson, Mark A. ; Shultz, Leonard D. ; Lagasse, Eric ; Greiner, Dale L. ; Kaestner, Klaus H. ; Grompe, Markus. / Generation of islet-like cells from mouse gall bladder by direct ex vivo reprogramming. In: Stem Cell Research. 2013 ; Vol. 11, No. 1. pp. 503-515.
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