title: Electrofusion of mesenchymal stem cells and islet cells for diabetes therapy: a rat model.
abstract: Islet transplantation is a minimally invasive treatment for severe diabetes. However, it often requires multiple donors to accomplish insulin-independence and the long-term results are not yet satisfying. Therefore, novel ways to overcome these problems have been explored. Isolated islets are fragile and susceptible to pro-apoptotic factors and poorly proliferative. In contrast, mesenchymal stem cells (MSCs) are highly proliferative, anti-apoptotic and pluripotent to differentiate toward various cell types, promote angiogenesis and modulate inflammation, thereby studied as an enhancer of islet function and engraftment. Electrofusion is an efficient method of cell fusion and nuclear reprogramming occurs in hybrid cells between different cell types. Therefore, we hypothesized that electrofusion between MSC and islet cells may yield robust islet cells for diabetes therapy. We establish a method of electrofusion between dispersed islet cells and MSCs in rats. The fusion cells maintained glucose-responsive insulin release for 20 days in vitro. Renal subcapsular transplantation of fusion cells prepared from suboptimal islet mass (1,000 islets) that did not correct hyperglycemia even if co-transplanted with MSCs, caused slow but consistent lowering of blood glucose with significant weight gain within the observation period in streptozotocin-induced diabetic rats. In the fusion cells between rat islet cells and mouse MSCs, RT-PCR showed new expression of both rat MSC-related genes and mouse β-cell-related genes, indicating bidirectional reprogramming of both β-cell and MSCs nuclei. Moreover, decreased caspase3 expression and new expression of Ki-67 in the islet cell nuclei suggested alleviated apoptosis and gain of proliferative capability, respectively. These results show that electrofusion between MSCs and islet cells yield special cells with β-cell function and robustness of MSCs and seems feasible for novel therapeutic strategy for diabetes mellitus.
authors.affiliation: Department of Organ Reconstruction, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
authors.forename: Goichi
authors.initials: G
authors.lastname: Yanai
authors.forename: Takashi
authors.initials: T
authors.lastname: Hayashi
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhi
authors.forename: Kai-Chiang
authors.initials: KC
authors.lastname: Yang
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Shirouzu
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimabukuro
authors.forename: Akihito
authors.initials: A
authors.lastname: Hiura
authors.forename: Kazutomo
authors.initials: K
authors.lastname: Inoue
authors.forename: Shoichiro
authors.initials: S
authors.lastname: Sumi
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis; D001786:Blood Glucose / Q000378:metabolism; D001835:Body Weight; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002459:Cell Fusion / Q000379:methods*; D049109:Cell Proliferation; D065150:Cellular Reprogramming; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000235:genetics / Q000473:pathology* / Q000628:therapy*; D004195:Disease Models, Animal; D004563:Electrochemistry / Q000379:methods*; D005453:Fluorescence; D005786:Gene Expression Regulation; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D016381:Islets of Langerhans Transplantation; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D051381:Rats; D015203:Reproducibility of Results; D013194:Staining and Labeling
chemical_list: D015703:Antigens, CD; D001786:Blood Glucose; D007328:Insulin; D053148:Caspase 3