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Cooperation by fibroblasts and bone marrow-mesenchymal stem cells to improve pancreatic rat-to-mouse islet xenotransplantation.

Perez-Basterrechea M, Obaya AJ, Meana A, Otero J, Esteban MM - PLoS ONE (2013)

Bottom Line: Histology of immunocompetent mice showed an attenuation of the immune response in those grafts with BM-MSCs, which was improved by co-transplantation with fibroblasts, since they increased BM-MSC survival.In summary, fibroblasts and BM-MSCs showed similar pro-angiogenic properties in this model of islet xenotransplantation, whereas only BM-MSCs exerted an immunomodulatory effect, which was improved by the presence of fibroblasts.These results suggest that cooperation of different cell types with islets will be required to achieve a long-term functional graft.

View Article: PubMed Central - PubMed

Affiliation: Transplants, Cell therapy and Regenerative Medicine Unit, Hospital Universitario Central de Asturias, Oviedo, Spain. marcos.perez@sespa.princast.es

ABSTRACT
Experimental and clinical experiences highlight the need to review some aspects of islet transplantation, especially with regard to site of grafting and control of the immune response. The subcutaneous space could be a good alternative to liver but its sparse vasculature is its main limitation. Induction of graft tolerance by using cells with immunoregulatory properties is a promising approach to avoid graft rejection. Both Fibroblasts and Mesenchymal Stem Cells (MSCs) have shown pro-angiogenic and immunomodulatory properties. Transplantation of islets into the subcutaneous space using plasma as scaffold and supplemented with fibroblasts and/or Bone Marrow-MSCs could be a promising strategy to achieve a functional extra-hepatic islet graft, without using immunosuppressive drugs. Xenogenic rat islets, autologous fibroblasts and/or allogenic BM-MSCs, were mixed with plasma, and coagulation was induced to constitute a Plasma-based Scaffold containing Islets (PSI), which was transplanted subcutaneously both in immunodeficient and immunocompetent diabetic mice. In immunodeficient diabetic mice, PSI itself allowed hyperglycemia reversion temporarily, but the presence of pro-angiogenic cells (fibroblasts or BM-MSCs) within PSI was necessary to improve graft re-vascularization and, thus, consistently maintain normoglycemia. In immunocompetent diabetic mice, only PSI containing BM-MSCs, but not those containing fibroblasts, normalized glycemia lasting up to one week after transplantation. Interestingly, when PSI contained both fibroblasts and BM-MSCs, the normoglycemia period showed an increase of 4-times with a physiological-like response in functional tests. Histology of immunocompetent mice showed an attenuation of the immune response in those grafts with BM-MSCs, which was improved by co-transplantation with fibroblasts, since they increased BM-MSC survival. In summary, fibroblasts and BM-MSCs showed similar pro-angiogenic properties in this model of islet xenotransplantation, whereas only BM-MSCs exerted an immunomodulatory effect, which was improved by the presence of fibroblasts. These results suggest that cooperation of different cell types with islets will be required to achieve a long-term functional graft.

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Histology of subcutaneous islet xenografts in immunocompetent diabetic mice.A) Representative images of subcutaneously transplanted islets alone (ISC), PSI without cells and PSI containing: 106 autologous fibroblasts (PSI-5F), 106 allogenic BM-MSCs (PSI-5M), or 106 of both autologous fibroblasts and allogenic BM-MSCs (PSI-5FM). Islet grafts were retrieved seven days after transplantation in diabetic mice. Samples were stained with H and E, or labelled with anti-insulin or anti-MPO antibodies. Black arrows point to islets. B) Quantification of the number of MPO-positive cells per field present on the leukocyte infiltration of the islet graft. C) Quantification of the insulin-positive area per section present on the islet graft. (**) p<0.01; (***) p<0.001.
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pone-0073526-g004: Histology of subcutaneous islet xenografts in immunocompetent diabetic mice.A) Representative images of subcutaneously transplanted islets alone (ISC), PSI without cells and PSI containing: 106 autologous fibroblasts (PSI-5F), 106 allogenic BM-MSCs (PSI-5M), or 106 of both autologous fibroblasts and allogenic BM-MSCs (PSI-5FM). Islet grafts were retrieved seven days after transplantation in diabetic mice. Samples were stained with H and E, or labelled with anti-insulin or anti-MPO antibodies. Black arrows point to islets. B) Quantification of the number of MPO-positive cells per field present on the leukocyte infiltration of the islet graft. C) Quantification of the insulin-positive area per section present on the islet graft. (**) p<0.01; (***) p<0.001.

Mentions: Histological analysis of subcutaneous islet xenografts was performed seven days after transplantation in diabetic immunocompetent mice (Figure 4). Xenografts of ISC, PSI without cells or containing autologous fibroblasts (PSI-5F) showed a great lymphocyte and polymorphonuclear cells (PMN) infiltration. No insulin-positive cells were observed in ISC grafts, while PSI and PSI-5F grafts showed small aggregates of insulin-positive cells, resulting from islet degradation, surrounded by non-integrated plasma scaffold. Xenografts containing allogenic BM-MSCs (PSI-5M) showed less lymphocyte infiltration with a lower number of PMNs. Some insulin-positive islets were observed, but most of them showed symptoms of injury like disrupted insulin staining and irregular morphology. Finally, xenografts containing autologous fibroblasts and allogenic BM-MSCs (PSI-5FM) showed the least lymphocyte and PMN infiltration. Furthermore, several insulin-positive islets were observed, most of them with normal morphology and continuous insulin staining.


Cooperation by fibroblasts and bone marrow-mesenchymal stem cells to improve pancreatic rat-to-mouse islet xenotransplantation.

Perez-Basterrechea M, Obaya AJ, Meana A, Otero J, Esteban MM - PLoS ONE (2013)

Histology of subcutaneous islet xenografts in immunocompetent diabetic mice.A) Representative images of subcutaneously transplanted islets alone (ISC), PSI without cells and PSI containing: 106 autologous fibroblasts (PSI-5F), 106 allogenic BM-MSCs (PSI-5M), or 106 of both autologous fibroblasts and allogenic BM-MSCs (PSI-5FM). Islet grafts were retrieved seven days after transplantation in diabetic mice. Samples were stained with H and E, or labelled with anti-insulin or anti-MPO antibodies. Black arrows point to islets. B) Quantification of the number of MPO-positive cells per field present on the leukocyte infiltration of the islet graft. C) Quantification of the insulin-positive area per section present on the islet graft. (**) p<0.01; (***) p<0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756982&req=5

pone-0073526-g004: Histology of subcutaneous islet xenografts in immunocompetent diabetic mice.A) Representative images of subcutaneously transplanted islets alone (ISC), PSI without cells and PSI containing: 106 autologous fibroblasts (PSI-5F), 106 allogenic BM-MSCs (PSI-5M), or 106 of both autologous fibroblasts and allogenic BM-MSCs (PSI-5FM). Islet grafts were retrieved seven days after transplantation in diabetic mice. Samples were stained with H and E, or labelled with anti-insulin or anti-MPO antibodies. Black arrows point to islets. B) Quantification of the number of MPO-positive cells per field present on the leukocyte infiltration of the islet graft. C) Quantification of the insulin-positive area per section present on the islet graft. (**) p<0.01; (***) p<0.001.
Mentions: Histological analysis of subcutaneous islet xenografts was performed seven days after transplantation in diabetic immunocompetent mice (Figure 4). Xenografts of ISC, PSI without cells or containing autologous fibroblasts (PSI-5F) showed a great lymphocyte and polymorphonuclear cells (PMN) infiltration. No insulin-positive cells were observed in ISC grafts, while PSI and PSI-5F grafts showed small aggregates of insulin-positive cells, resulting from islet degradation, surrounded by non-integrated plasma scaffold. Xenografts containing allogenic BM-MSCs (PSI-5M) showed less lymphocyte infiltration with a lower number of PMNs. Some insulin-positive islets were observed, but most of them showed symptoms of injury like disrupted insulin staining and irregular morphology. Finally, xenografts containing autologous fibroblasts and allogenic BM-MSCs (PSI-5FM) showed the least lymphocyte and PMN infiltration. Furthermore, several insulin-positive islets were observed, most of them with normal morphology and continuous insulin staining.

Bottom Line: Histology of immunocompetent mice showed an attenuation of the immune response in those grafts with BM-MSCs, which was improved by co-transplantation with fibroblasts, since they increased BM-MSC survival.In summary, fibroblasts and BM-MSCs showed similar pro-angiogenic properties in this model of islet xenotransplantation, whereas only BM-MSCs exerted an immunomodulatory effect, which was improved by the presence of fibroblasts.These results suggest that cooperation of different cell types with islets will be required to achieve a long-term functional graft.

View Article: PubMed Central - PubMed

Affiliation: Transplants, Cell therapy and Regenerative Medicine Unit, Hospital Universitario Central de Asturias, Oviedo, Spain. marcos.perez@sespa.princast.es

ABSTRACT
Experimental and clinical experiences highlight the need to review some aspects of islet transplantation, especially with regard to site of grafting and control of the immune response. The subcutaneous space could be a good alternative to liver but its sparse vasculature is its main limitation. Induction of graft tolerance by using cells with immunoregulatory properties is a promising approach to avoid graft rejection. Both Fibroblasts and Mesenchymal Stem Cells (MSCs) have shown pro-angiogenic and immunomodulatory properties. Transplantation of islets into the subcutaneous space using plasma as scaffold and supplemented with fibroblasts and/or Bone Marrow-MSCs could be a promising strategy to achieve a functional extra-hepatic islet graft, without using immunosuppressive drugs. Xenogenic rat islets, autologous fibroblasts and/or allogenic BM-MSCs, were mixed with plasma, and coagulation was induced to constitute a Plasma-based Scaffold containing Islets (PSI), which was transplanted subcutaneously both in immunodeficient and immunocompetent diabetic mice. In immunodeficient diabetic mice, PSI itself allowed hyperglycemia reversion temporarily, but the presence of pro-angiogenic cells (fibroblasts or BM-MSCs) within PSI was necessary to improve graft re-vascularization and, thus, consistently maintain normoglycemia. In immunocompetent diabetic mice, only PSI containing BM-MSCs, but not those containing fibroblasts, normalized glycemia lasting up to one week after transplantation. Interestingly, when PSI contained both fibroblasts and BM-MSCs, the normoglycemia period showed an increase of 4-times with a physiological-like response in functional tests. Histology of immunocompetent mice showed an attenuation of the immune response in those grafts with BM-MSCs, which was improved by co-transplantation with fibroblasts, since they increased BM-MSC survival. In summary, fibroblasts and BM-MSCs showed similar pro-angiogenic properties in this model of islet xenotransplantation, whereas only BM-MSCs exerted an immunomodulatory effect, which was improved by the presence of fibroblasts. These results suggest that cooperation of different cell types with islets will be required to achieve a long-term functional graft.

Show MeSH
Related in: MedlinePlus