Plastic Surgery Research Council
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Presenter: Damon S Cooney, MD
Co-Authors: Wu LW; Sarkar K; Mao Q; Lee WP; Brandacher G; Cooney DS; Yuan N
Johns Hopkins University School of Medicine

Background: Adipose-derived stem cells (ASCs) are promising sources of immunomodulation for hand and face transplantation, but their effects have not yet been investigated in a murine model of reconstructive transplantation. The relative effects of donor- vs. recipient-derived ASCs have also not been studied.

Methods: Plastic-adherent ASCs isolated from C57B6 and Balb/c mice were cultured in high-glucose Dulbecco s Modified Eagle Medium, 10% FBS, and 1% penicillin-streptomycin and used at passages 2-5. Flow cytometry was performed to identify surface markers consistent with the ASC phenotype. CFSE-stained C57B6 splenocytes were cultured in a 1:1 ratio with mitomycin-treated Balb/c splenocytes (allogeneic stimulation) or with mitomycin-treated, non-CFSE-stained C57B6 splenocytes (control). Stimulated and control wells were cultured with varying doses of ASCs from C57B6 and Balb/c origin. Samples were harvested at day 4, incubated with anti-mouse CD3 antibody, and analyzed for proliferation via flow cytometry.

Results: Cultured cells were rich in CD105 and CD29 and negative for CD11b, CD34, and CD45. Allogeneically stimulated wells showed significant cell proliferation as compared to autologous controls. Murine ASCs dramatically suppressed proliferation of allogeneically stimulated CD3-positive lymphocytes. Effects were dose-dependent and potent; even a ratio of 1000:1 ASCs showed some immunoregulatory effect. Similar doses of C57B6 ASCs and Balb/c ASCs showed no difference in effects.

Conclusion: We demonstrate potent immunomodulation by ASCs in an in vitro mouse model of allo-stimulation, confirming for the first time the feasibility of studying ASCs in mouse models of reconstructive transplantation. The mouse model is particularly valuable for transplantation research, given the availability of mouse-specific reagents and genetically modified animals that can elucidate biological mechanisms. We observed no difference between ASCs derived from donor and recipient sources. This work prepares us for upcoming studies of the in vivo effects of ASCs.

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