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Theme: Inclusion and Collaboration Theme: Inclusion and Collaboration

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Muscle Regeneration and Function Restoration Following Transplantation of Chimeric Cells of Myoblast And Mesenchymal Stem Cell Origin
Erzsebet Szilagyi, MD, PhD; Joanna Cwykiel, MSc; Anna Domaszewska-Szostek, PhD; Maria Siemionow, MD PhD DSc
University of Illinois at Chicago, Chicago, IL

Background: Allogeneic stem cell therapies aim to restore muscle tissue after traumatic loss or muscular dystrophies. Limited engraftment due to allogeneic rejection is a major challenge. Muscle Derived Chimeric Cells (MDCC), created via ex vivo fusion of myoblast (MB) and mesenchymal stem cells (MSC), represent a promising therapeutic option. The aim of this study was to characterize human MDCC in vitro and to assess MDCC efficacy in engraftment and restoration of muscle function in Duchenne Muscular Dystrophy (DMD) mdx/scid mice model.

Methods: MDCC proliferation, dystrophin expression and myogenic differentiation was tested. Lymphocyte proliferation assay was performed to evaluate MDCC immunogenicity. To test efficacy of MDCC in vivo, mdx/scid mice received intramuscular injections to gastrocnemius muscle (GM): Group 1 - vehicle (60mcl PBS), Group 2 - 0.25x106 of MSC and 0.25x106 MB, Group 3 - 0.5x106 of MB/MSC MDCC. Therapeutic effect was monitored by muscle function tests. DE was evaluated at day 7 and 90 after MDCC delivery.

Results: Parent cells genotype, dystrophin expression, proliferation and differentiation to mature skeletal myocytes of MDCC was confirmed. Proliferation of responder lymphocytes after stimulation with MDCC was 5.49% (SI=3.15), compared to positive controls of 55% (SI=31) after stimulation with 3rd party lymphocyte and 10% (SI=5.8) after stimulation with MB parent cells. MDCC survival and engraftment to GM was confirmed by dystrophin expression of 17.7% at day 7 and 20.26% at 90 days . Moreover, dystrophin, HLA-ABC and mature myocyte-specific markers were co-expressed by MDCC confirming human origin and differentiation of MDCC after transplant. MDCC recipients showed increase in muscle force (p=0.04) and improved fatigue tolerance compared to vehicle group.

Conclusion: This study confirmed efficacy of MDCC therapy in restoration of muscle function, which correlated with dystrophin expression in the GM of mdx/scid mice. MDCC therapy represents a novel, universal approach for restoration of muscle function in muscular dystrophy, traumatic loss and for regeneration of muscle components of VCA.

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