Recipient Adipose-derived Stem Cells Enhance Recipient Cell Engraftment and Prolong Allotransplant Survival in a Miniature Swine Hind-limb Model

Abstract

Donor mesenchymal stem cells (MSCs) could prolong vascularized composite allotransplantation (VCA) survival in our previous studies. However, recipient adipose tissue is easier to harvest than donor tissue for preconditioning modulation. Hence, this study investigated the efficacy of recipient autologous adipose-derived stem cells (rADSCs) for VCA survival. The heterotopic hind-limb transplantation from female donor to male recipient was performed in outbred miniature swine. Group I (n ¼ 6) was untreated controls. Group II (n ¼ 4) obtained rADSCs infusions (given on weeks 0, þ1, þ2, and þ3). Group III (n ¼ 4) obtained tacrolimus (FK506, weeks 0 to þ4). Group IV (n ¼ 8) received irradiation (IR; day À1), FK506 (weeks 0 to þ4), and rADSC infusions (weeks 0, þ1, þ2, and þ3). The results revealed treatment with multiple injections of rADSCs along with IR and FK506 resulted in a statistically significant increase in allograft survival. The percentage of CD4þ regulatory T cells were significantly increased in the rADSC-IR-FK506 group as compared to controls. Analysis of recipient peripheral blood revealed that transforming growth factor b1 (TGFb1) was significantly increased in the rADSC-IR-FK506 group. The polymerase chain reaction (PCR) analysis and immunohistochemical staining showed recipient sex-determining region of Y (SRY) chromosome gene expression existed in donor allotissues in the rADSC-IR-FK506 group. These results indicate that rADSCs in addition to IR and transient immunosuppressant could prolong allotransplant survival, modulate T-cell regulation, and enhance recipient cell engraftment into the allotransplant tissues.

Clinical and preclinical tolerance protocols for vascularized composite allograft transplantation

“…Adipose-derived stem cells are harvested from the recipient and require no pre-conditioning, therefore making their use less taxing to the recipient 64 . In adipose-derived stem cell studies, Kou et al 64 found that outbred swine treated with adiposederived stem cells, tacrolimus, and irradiation (n = 8) had significantly increased graft survival over 196 days. They also found an increase in regulatory T-cells in this group at 6 weeks posttransplant, but these levels normalized by week 15.…”

Research Status of Mesenchymal Stem Cells in Liver Transplantation

“…For CNIs, MSCs combined with tacrolimus demonstrated considerable coordination in immune regulation after organ transplantation. Kuo et al found that MSCs plus tacrolimus improved graft survival in a miniature swine hind-limb model 124 . Suda and his group also showed that the combination of MSCs and tacrolimus could relieve ischemic brain damage in rats 125 .…”

Harvest and Culture of Porcine Adipose-Derived Mesenchymal Stem Cells for Autologous Transplantation

“…Several papers have briefly discussed porcine fat isolation and characterization but only as part of an in vivo experiment or in experiments without survival surgeries. Many of these studies have used culture conditions identical to those used for humans [3][4][5][6][7][8][9][10][11][12][13]. Here, we provide specific surgical techniques for porcine fat harvest as a survival surgery and describe pADMSC isolation and culture optimization for autologous transplantation, including culture conditions that produce porcine MSCs that are analogous to those cultured from humans, with similar proliferation and differentiation capacities.…”