title: β-MSCs: successful fusion of MSCs with β-cells results in a β-cell like phenotype.
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.
authors.affiliation: Centre for Biomolecular Interactions, University of Bremen, Bremen, Germany.|Department of Cell and Gene Therapy, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Azizi
authors.affiliation: Department of Cell and Gene Therapy, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Lange
authors.affiliation: Centre for Biomolecular Interactions, University of Bremen, Bremen, Germany.
authors.forename: Federico
authors.initials: F
authors.lastname: Paroni
authors.affiliation: Centre for Biomolecular Interactions, University of Bremen, Bremen, Germany.
authors.forename: Amin
authors.initials: A
authors.lastname: Ardestani
authors.affiliation: Centre for Biomolecular Interactions, University of Bremen, Bremen, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Meyer
authors.affiliation: Centre for Biomolecular Interactions, University of Bremen, Bremen, Germany.|Department of Laboratory Medicine, Peking University Third Hospital, Beijing, China.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Cell and Gene Therapy, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Axel R
authors.initials: AR
authors.lastname: Zander
authors.affiliation: Departments of Medicine and Physiology, University of Utah and George E Wahlen Department of Veterans Affairs Medical Center, Salt Lake City, Utah, USA.
authors.forename: Christof
authors.initials: C
authors.lastname: Westenfelder
authors.affiliation: Centre for Biomolecular Interactions, University of Bremen, Bremen, Germany.|German Center for Diabetes Research (DZD) project partner, University of Bremen, Bremen, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Maedler
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D002459:Cell Fusion / Q000379:methods*; D002469:Cell Separation; D018920:Coculture Techniques; D005260:Female; D005947:Glucose / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D006801:Humans; D007328:Insulin / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009687:Nuclear Proteins; D010641:Phenotype; D010835:Phytohemagglutinins / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D051381:Rats; D014157:Transcription Factors
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005947:Glucose; D049452:Green Fluorescent Proteins; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D007328:Insulin; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D010835:Phytohemagglutinins; D011092:Polyethylene Glycols; D014157:Transcription Factors; C000715631:NKX2-2 protein, human; C000715628:Nkx2-2 protein, rat; C093564:Neurogenic differentiation factor 1
keywords: MSCs; Pathology Section; beta-cells; cell fusion; diabetes; islets