Reprogramming mouse embryo fibroblasts to functional islets without genetic manipulation

Chandravanshi, Bhawna and Bhonde, Ramesh R (2018) Reprogramming mouse embryo fibroblasts to functional islets without genetic manipulation. Journal of Cellular Physiology, 233 (2). pp. 1627-1637. ISSN 1097-4652

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Abstract

Mouse embryo fibroblast cells isolated from 12-14 day old pregnant mice were characterized for their surface markers and tri-lineage differentiation potential. They were subjected to serum free media containing a cocktail of islet differentiating reagents and analysed for the expression of pancreatic lineage transcripts. The islet like cell aggregates (ICAs) was confirmed for their islet like properties via immunofluorecence for C-peptide, glucagon and somatostain. They were positive for CD markers-Sca1, CD44, CD73 and CD90 and negative for haematopoietic markers- CD34 and CD45 at both transcription and translational levels. The transcriptional analysis of the ICAs at different day points exhibited up-regulation of islet markers (Insulin, PDX1, HNF3, Glucagon and Somatostatin) and down-regulation of MSC- markers (Vimentin and Nestin). They positively stained for dithizone, C-peptide, insulin, glucagon and somatostatin indicating intact insulin producing machinery. In-vitro glucose stimulation assay revealed 3 fold increase in insulin secretion as compared to basal glucose with insulin content being the same in both the conditions. The preliminary in vivo data on ICA transplantation showed reversal of diabetes in streptozotocin induced diabetic mice.

Item Type: Article
Uncontrolled Keywords: Mouse embryo fibroblast; mesenchymal stem cells; insulin like cell aggregates; dithizone; C-peptide.
Subjects: Regenerative Medicine > MIRM Bangalore
Depositing User: MCON Library
Date Deposited: 12 Nov 2017 04:15
Last Modified: 12 Nov 2017 04:15
URI: http://eprints.manipal.edu/id/eprint/149959

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