Efficient Differentiation of AR42J Cells towards Insulin-Producing Cells using Pancreatic Transcription Factors in Combination with Growth Factors

Maria Joao Lima, Hilary Docherty, Y. Chen, Kevin Docherty

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

The AR42J-B13 rat pancreatic acinar cell line was used to identify pancreatic transcription factors and exogenous growth factors (GFs) that might facilitate the reprogramming of exocrine cells into islets. Adenoviruses were used employed to induce exogenous expression of the pancreatic transcription factors (TFs) Pdx1, MafA, Ngn3 and Pax4. Individually Pdx1, MafA and Pax4 had no effect on the expression of endocrine markers, whilst adeno-Ngn3 on its own increased the expression of Pax4, Ngn3 and NeuroD. In combination the four TFs had a significant effect on the expression of insulin 1 and insulin 2 that was associated with a change in cell morphology from a rounded to a spindle-like shape. Amongst a range of growth factors, Betacellulin and Nicotinamide were shown to enhance the effects of the four TFs. The presence of adeno-Pax4 in the differentiation cocktail was important in limiting the expression of glucagon and in generating glucose sensitive insulin secretion. Further experiments asked whether the adenoviral TFs could be replaced by protein transduction domain (PTD)-containing TFs. The results showed that the PTD-TFs could mimic in part the effects of the adeno-TFs, but the resultant cells did not undergo the important morphological change associated with differentiation to endocrine lineages and levels of endogenous markers were very much lower. In summary, the results describe a cocktail of four TFs and two GFs that can be used to induce formation of glucose sensitive insulin secreting cells from ARJ42 cells, and demonstrate that it would be difficult to replace adenoviral transduction with PTD-TFS.
Original languageEnglish
Pages (from-to)69-80
Number of pages12
JournalMolecular and Cellular Endocrinology
Volume358
Issue number1
Early online date10 Mar 2012
DOIs
Publication statusPublished - 6 Jul 2012

Fingerprint

Cell Differentiation
Intercellular Signaling Peptides and Proteins
Transcription Factors
Cells
Insulin
Glucose
Proteins
Niacinamide
Acinar Cells
Insulin-Secreting Cells
Glucagon
Islets of Langerhans
Adenoviridae
Rats
Cell Line
Protein Domains

Keywords

  • stem cells
  • diabetes
  • islets of langerhans
  • endocrine pancreas
  • Pdx1

Cite this

Efficient Differentiation of AR42J Cells towards Insulin-Producing Cells using Pancreatic Transcription Factors in Combination with Growth Factors. / Lima, Maria Joao; Docherty, Hilary; Chen, Y.; Docherty, Kevin.

In: Molecular and Cellular Endocrinology, Vol. 358, No. 1, 06.07.2012, p. 69-80.

Research output: Contribution to journalArticle

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