Regulation Of Human Adipose And Umbilical Cord Mesenchymal Stem Cell On Mice Allogeneic Corneal Transplantation

Purpose:Mesenchymal stem cell is a heterogeneous population of non-hematopoietic pluripotent cell that exert a broad range of immunosuppressive functions in a variety of diseases.The role and functional differences of human-derived MSC have been less studied in the field of corneal transplantation.In this study,we established the mice model of corneal allotransplantation to investigate the potential of systemic or subconjunctival application of human umbilical cord mesenchymal stem cell or human adipose mesenchymal stem cells to promote the survival of corneal allografts in mice and to investigate the effects of human umbilical cord mesenchymal stem cell or human adipose mesenchymal stem cell on grafts angiogenesis and lymphangiogenesis.Methods:MSC of different sources were isolated and extracted from human adipose and umbilical cord tissues.The cells were identified by observation of their morphology during culture,flow cytometry detection of cell surface markers,and induction of osteogenic,lipogenic,and chondrogenic differentiation results.C57BL/6 mice were used as the donor and BALB/c mice as the recipient to establish the model of allograft penetrating keratoplasty in mice.On the day of transplantation,10~6 hUC-MSC or hAD-MSC were injected intravenously behind the globe,or a subconjunctival injection of 5x10~5 hUC-MSC or hAD-MSC on the day of transplantation.The corneal grafts were observed and photographed with a slit lamp every other day after surgery,and the survival of the grafts was judged and continued for 45 days.In addition,15 days after surgery,partial corneal grafts were collected for H&E staining to evaluate pathological changes,and immunofluorescence staining was used to detect the angiogenesis and lymphangiogenesis of the corneal grafts,and quantitative analysis was performed.Results:Cell identification results showed that purified,well-performing MSC could be stably obtained from human adipose and human umbilical cord tissues.Both hAD-MSC and hUC-MSC injected by different routes significantly prolonged the survival time of the grafts compared to the control group.Pathological results showed greatly improved corneal graft turbidity,collagen fiber disorder,interstitial edema,and inflammatory cell infiltration.Immunofluorescence staining results showed a reduction in the area of corneal graft neovascularization and lymphatic vessel perfusion.Conclusions:Systemic or subconjunctival application of hAD-MSC or hUC-MSC induces corneal immune tolerance formation,promotes long-term graft survival,and inhibits corneal angiogenesis and lymphangiogenesis in the early post-transplant period.