Induced pluripotent stem cell‐derived mesenchymal stem cells prolong hind limb survival in a rat vascularized composite allotransplantation model

Abstract

Background The reduction of systemic immunosuppressive agents is essential for the expansion of vascularized composite allotransplantation (VCA) in a clinical setting. The purpose of this study is to compare human‐induced pluripotent stem cell‐derived mesenchymal stem cells (iMSCs) with four other types of mesenchymal stem cells (human bone marrow‐derived MSCs [BMMSCs], human adipose‐derived MSCs [ADMSCs], rat BMMSCs, and rat ADMSCs) in vitro, and to investigate the in vivo immunomodulatory effect of iMSCs in a rat VCA model. Materials and methods One Brown Norway (BN) rat, 2 Lewis (LEW) rats, and 1 Wistar rat were used in the mixed lymphocyte reaction (MLR), and 9 BN rats and 3 LEW rats (for donors), and 24 LEW rats (for recipients) were used in the VCA model. The abovementioned five types of MSCs were imaged to examine their morphology and were also tested for suppressor function using a MLR. The 24 recipient LEW rats were divided randomly into four groups, and subjected to orthotopic hind limb transplantation. The three control groups were the Iso group, in which transplantation was performed on from three to six LEW rats without immunosuppressive treatment (n = 6); the FK group, in which transplantation was performed from BN rats to LEW rats and recipient rats were treated with tacrolimus alone (FK 506, 0.2 mg/kg, days 0–6 postoperatively, intraperitoneally) (n = 6); and the UT group, in which transplantation was performed from BN rats to LEW rats without any immunosuppressive treatment (n = 6). The experimental group was the iMSC group, in which transplantation was performed from BN rats to LEW rats and recipient rats were treated with tacrolimus (FK 506, 0.2 mg/kg, days 0–6 postoperatively, intraperitoneally) and injected with iMSCs (2 × 106 cells, day 7, intravenously) (n = 6). Hind limb survival was assessed by daily inspection of gross appearance until 50 days postoperatively. Histology of the skin and muscle biopsy were investigated on day 14 postoperatively. A time series of the plasma cytokine level (before transplantation, and at 10, 14, and 17 days after transplantation) was also analyzed. Results The size of adherent and trypsinized iMSCs was 67.5 ± 8.7 and 9.5 ± 1.1 μm, respectively, which was the smallest among the five types of MSCs (p < .01). The absorbance in MLR was significantly smaller with rat ADMSCs (p = .0001), human iMSCs (p = .0006), rat BMMSCs (p = .0014), human ADMSCs (p = .0039), and human BMMSCs (p = .1191) compared to without MSCs. In vivo, iMSC treatment prolonged hind limb survival up to 12.7 days in macroscopic appearance, which is significantly longer than that of the FK group (p < .01). Histology of the skin and muscle biopsy revealed that mononuclear cell infiltration was significantly reduced by iMSC injection (p < .01). iMSC treatment also affected proinflammatory cytokines (interferon‐gamma (IFNγ) and tumor necrosis factor α (TNFα)) and the anti‐inflammatory cytokine (interleukin‐10 (IL‐10)) of the recipient plasma. The IFNγ levels at Δ14 and the TNFα levels at Δ...

In Vivo Regeneration of Rat Laryngeal Cartilage with Mesenchymal Stem Cells Derived from Human Induced Pluripotent Stem Cells Via Neural Crest Cells

“…X-SCID rats were utilized to avoid immune rejection by the host; however, the immune systems of these rats are not completely depleted (Mashimo et al, 2010). It is known that human MSCs have immunomodulatory properties (Madrigal et al, 2014), and it has been demonstrated that iMSCs induced from iNCCs also have immunomodulatory properties under xenogeneic conditions (Mitsuzawa et al, 2019). Allogeneic MSC transplantation has been widely used in clinical settings, and HLAediting techniques may provide safety and feasibility in using hiPSCs for allogeneic transplantation (Xu et al, 2019).…”

Cell therapy in vascularized composite allotransplantation

“…It has been hypothesized that BM-MSCs may prolong VCA survival through the increased levels of CD4 + CD25 + Tregs, TGF-β1, and serum IL-10 [ 70 ]. A recent study illustrated the induced pluripotent stem cell-derived mesenchymal stem cells (MSCs) also prolonged VCA allograft survival through the induction of T cell hyporesponsiveness, reduction of proinflammatory cytokines (interferon gamma and tumor necrosis factor α), and elevation of the anti-inflammatory cytokine, IL-10 [ 71 ]. AD-MSCs also prolonged VCA allograft survival by suppressing effector T cells and enhancing Tregs [ Table 2 ] [ 72 , 73 ].…”

“…(2014) [ 81 ] Donor HSCs Swine Fascio-cutaneous 15 × 10 9 (0), IV T cell depletion + non-myeloablatıve TBI + CsA (0–45) Prolong allograft 85–100 days Induce peripheral blood chimerism between 20 and 100% Mitsuzawa et al. (2019) [ 71 ] Syngeneic iMSCs Rat Hindlimb 2 × 10 6 (7), IV Tac (0–6) Prolong allografts >17–21 days Induce T cell donor-specific hyporesponsiveness Kuo et al. (2017) [ 31 ] Autologous AD-MSCs Swine Hindlimb 1 × 10 6 /kg (0–3), IV TBI (−1) + Tac (0–4) Prolong allografts >196 days Modulate T cell regulation Plock et al.…”

Application of mesenchymal stem cells derived from human pluripotent stem cells in regenerative medicine

“…iPSC-MSCs also suppressed T-cell effector cells of Th1/Th2 and increased regulatory T cell (Treg) response[ 105 ]. iPSC-MSCs prolonged hind limb survival by reducing mononuclear cell infiltration, lowering TNFα and IFNγ, increasing interleukin 10, and thus protecting against acute rejection in a rat vascularized composite allotransplantation model[ 106 ] (Figure 1 ). iPSC-MSCs disrupted NK cell cytolytic machinery to prevent allograft rejection by decreasing activation markers and ERK1/2 signalling, leading to impaired immunologic synapses and secreted cytotoxic granules.…”