Genetically Modified Human Bone Marrow Derived Mesenchymal Stem Cells for Improving the Outcome of Human Islet Transplantation

Abstract

The objective of this study was to determine the potential of human bone marrow derived mesenchymal stem cells (hBMSCs) as gene carriers for improving the outcome of human islet transplantation. hBMSCs were characterized for the expression of phenotypic markers and transduced with Adv-hVEGF-hIL-1Ra to overexpress human vascular endothelial growth factor (hVEGF) and human interleukin-1 receptor antagonist (hIL-1Ra). Human islets were co-cultured with hBMSCs overexpressing hVEGF and hIL-1Ra. Islet viability was determined by membrane fluorescent method and glucose stimulation test. Transduced hBMSCs and human islets were co-transplanted under the kidney capsule of NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) diabetic mice and blood glucose levels were measured over time to demonstrate the efficacy of genetically modified hBMSCs. At the end of study, immunofluorescent staining of kidney section bearing islets was performed for insulin and von Willebrand Factor (vWF). hBMSCs were positive for the expression of CD73, CD90, CD105, CD146 and Stro-1 surface markers as determined by flow cytometry. Transduction of hBMSCs with adenovirus did not affect their stemness and differentiation potential as confirmed by mRNA levels of stem cell markers and adipogenic differentiation of transduced hBMSCs. hBMSCs were efficiently transduced with Adv-hVEGF-hIL-1Ra to overexpress hVEGF and hIL-1Ra. Live dead cell staining and glucose stimulation test have shown that transduced hBMSCs improved the viability of islets against cytokine cocktail. Co-transplantation of human islets with genetically modified hBMSCs improved the glycemic control of diabetic NSG mice as determined by mean blood glucose levels and intraperitoneal glucose tolerance test. Immunofluorescent staining of kidney sections was positive for human insulin and vWF. In conclusion, our results have demonstrated that hBMSCs may be used as gene carriers and nursing cells to improve the outcome of islet transplantation.

In VitroInduction of Human Adipose-Derived Stem Cells into Lymphatic Endothelial-Like Cells

“…1B). This pattern of expression was similar to previously characterized BM-MSCs, indicating that these two populations play similar roles in the body (Mundra et al, 2013;Tsai et al, 2014).…”

JAM-A promotes wound healing by enhancing both homing and secretory activities of mesenchymal stem cells

“…Intravenous injection of MSCs or gene-modified MSCs is an acceptable method for clinical application. As MSCs were expanded by demonstrating their capacity for self-renewal and multilineage differentiation, and gene modification can promote migratory ability, directional differentiation and secretion ability, etc., these characteristics are good for various tissue injuries [26,27]. Of course, MSC or JAM-A/MSC treatment depends on the type of wound, and the health status of the patient.…”

“…Various active factors endogenously secreted by MSCs play important roles in helping MSCs to repair affected tissues and organs by regulating the extracellular microenvironment, such as IFN-β and vascular endothelial growth factor (VEGF) [26]. However, the above genes could act as vectors to transport the target gene to the wound surface without enhancing the secretory function of MSCs [27].…”

Immunological Challenges Facing Translation of Alginate Encapsulated Porcine Islet Xenotransplantation to Human Clinical Trials

“…The results of studies utilizing autologous MSC transplantation also support the immunosuppressive capabilities of MSCs when co-transplanted with allogenic islets as evidenced by extension of the duration of normoglycemia in the recipient [ 105 -107 ]. However, in an attempt restoration of standard insulin secretion in response to plasma glucose levels for type 1 diabetics via xenogeneic transplants (islets co-transplanted with MSCs), current investigators have begun to focus on genetic modifi cation of porcine MSCs in hopes of achieving cross-species compatibility, and functionality [ 108 ].…”