Therapeutic Potential of Human-Induced Pluripotent Stem Cell-Derived Endothelial Cells in a Bleomycin-Induced SclerodermaMouse Model

Background and Aim: Vascular injury and destruction of endothelial cells(ECs) are the early events in scleroderma (SSc) patients. This study aims toinvestigate the therapeutic potential of human-induced pluripotent stem cell-derived ECs (hiPSC-ECs) to treat SSc. We have assessed the functionaldifferentiation of hiPSC-ECs and compared them with human embryonicstem cell-derived ECs (hESC-ECs) by a variety of in vitro experimentalapproaches. Additionally, we evaluated the therapeutic potential ofhiPSC-ECs in a bleomycin-induced SSc mouse model.Methods: The hESCs and hiPSCs culture and EC differentiation Humanumbilical vein endothelial cell isolation and culture Flow cytometryand cell sorting Immunostaining Colony-forming assay Uptake ofacetylated low-density lipoprotein (DiI-ac-LDL)Tube formation byECs in vitro Scleroderma mouse model and hiPSC-EC transplantationHistopathology and immunohistochemistry Mast cell number Enzymelinkedimmunosorbent assay of serum cytokineResults: Our results demonstrated that hiPSC-ECs and hESC-ECs showedsimilar maximum expressions of FLK1 (early EC marker) at day fiveduring differentiation. After sorting and culturing, the FLK1-positive cellsexhibited spindle and subsequent endothelial cobblestone morphologyin the EGM2 medium. The hESC-ECs and hiPSC-ECs also expressed lateEC markers CD31 (68% and 75%), CD144 (50% and 61%), CD146(46% and 61%), and DiI-labeled acetylated low-density lipoprotein (DiIac-LDL) uptake (55% and 63%), respectively. They additionally formedcapillary-like structures on Matrigel. Analyses of the transplantationof sorted CD31-positive hiPSC-ECs into the bleomycin-induced SScmouse model demonstrated that these cells participate in the recoveryof the damaged vessels. There was a reduction in collagen content; thenumber of total and degranulated mast cells returned to their normalstate, and bleomycin-induced wounds, as well as skin fibrosis, improvedfour weeks after transplantation of hiPSC-ECs. Our findings have shownthat the differentiation process fromhESCs and hiPSCs to vascular cellcomponents is similar. Besides, this is the first study to determine thetherapeutic potential of vascular cells from hiPSCs in the treatment of anSSc model. In the future, with further validation, these may be used as anappropriate source for the treatment of SSc patients.Conclusion: These early data have shown the proof of concept that thetransplantation of ECs derived from hiPSCs, similar to hESC-ECs, canprovide some benefits in the setting for SSc regeneration. With furthervalidation, the ECs that are isolated from hESCs and hiPSCs, as describedhere, can provide a continual endothelial cell source of therapy forvascular regeneration in SSc in addition to other diseases with endothelialabnormalities.

کلیدواژه ها:

hiPSCs ، Endothelial cells ، Differentiation ، Scleroderma disease ، Bleomycin animal model cell therapy