Oncotarget

Research Papers: Pathology:

β-MSCs: successful fusion of MSCs with β-cells results in a β-cell like phenotype

Zahra Azizi _, Claudia Lange, Federico Paroni, Amin Ardestani, Anke Meyer, Yonghua Wu, Axel R. Zander, Christof Westenfelder and Kathrin Maedler

PDF  |  HTML  |  Supplementary Files  |  How to cite  |  Order a Reprint

Oncotarget. 2016; 7:48963-48977. https://doi.org/10.18632/oncotarget.10214

Metrics: PDF 1869 views  |   HTML 2298 views  |   ?  


Abstract

Zahra Azizi1,2, Claudia Lange2, Federico Paroni1, Amin Ardestani1, Anke Meyer1, Yonghua Wu1,6, Axel R. Zander2, Christof Westenfelder3,4,* and Kathrin Maedler1,5,*

1 Centre for Biomolecular Interactions, University of Bremen, Bremen, Germany

2 Department of Cell and Gene Therapy, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

3 Departments of Medicine and Physiology, University of Utah, Salt Lake City, Utah, USA

4 George E Wahlen Department of Veterans Affairs Medical Center, Salt Lake City, Utah, USA

5 German Center for Diabetes Research (DZD) Project Partner, University of Bremen, Bremen, Germany

6 Department of Laboratory Medicine, Peking University Third Hospital, Beijing, China

* These authors have contributed equally to this work

Correspondence to:

Zahra Azizi, email:

Kathrin Maedler, email:

Christof Westenfelder, email:

Keywords: MSCs, cell fusion, beta-cells, islets, diabetes, Pathology Section

Received: October 26, 2015 Accepted: April 19, 2016 Published: June 21, 2016

Abstract

Bone marrow mesenchymal stromal cells (MSC) have anti-inflammatory, anti-apoptotic and immunosuppressive properties and are a potent source for cell therapy. Cell fusion has been proposed for rapid generation of functional new reprogrammed cells. In this study, we aimed to establish a fusion protocol of bone marrow−derived human MSCs with the rat beta-cell line (INS-1E) as well as human isolated pancreatic islets in order to generate insulin producing beta-MSCs as a cell-based treatment for diabetes.

Human eGFP+ puromycin+ MSCs were co-cultured with either stably mCherry-expressing rat INS-1E cells or human dispersed islet cells and treated with phytohemagglutinin (PHA-P) and polyethylene glycol (PEG) to induce fusion. MSCs and fused cells were selected by puromycin treatment.

With an improved fusion protocol, 29.8 ± 2.9% of all MSCs were β-MSC heterokaryons based on double positivity for mCherry and eGFP.

After fusion and puromycin selection, human NKX6.1 and insulin as well as rat Neurod1, Nkx2.2, MafA, Pdx1 and Ins1 mRNA were highly elevated in fused human MSC/INS-1E cells, compared to the mixed control population. Such induction of beta-cell markers was confirmed in fused human MSC/human dispersed islet cells, which showed elevated NEUROD1, NKX2.2, MAFA, PDX1 and insulin mRNA compared to the mixed control. Fused cells had higher insulin content and improved insulin secretion compared to the mixed control and insulin positive beta-MSCs also expressed nuclear PDX1. We established a protocol for fusion of human MSCs and beta cells, which resulted in a beta cell like phenotype. This could be a novel tool for cell-based therapies of diabetes.


Creative Commons License All site content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 License.
PII: 10214