In vivo tracking of the rejection and survival of mouse ovarian iso- and allo-grafts by bioluminescence imaging (BLI)

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OBJECTIVE: In this study, we utilized BLI as the real-time in vivo imaging to detect the fate-map of the iso- and allo-ovarian grafts. DESIGN: Transgenic mouse model. MATERIALS AND METHODS: The 8-week-old donors of FVB/N-Tg(PolII-luc)Ltc transgenic mice with luciferase gene fragment as a reporter are generated by pronuclei microinjection of PolII-Luc transgene into FVB/N fertilized embryos. Recipients (n=5 in each group) included group 1: FVB/N wild type (isogenic) with daily cyclosporine-A (CSA)(25 mg/kg) treatment, group 2: FVB/N wild type without CSA treatment, group 3: Balb/c wild type (allogenic) with daily CSA (25 mg/kg) treatment, group 4: Balb/c wild type without CSA treatment, and group 5: NOD.CB17-Prkdcscid/J (allogenic, immunoinsufficiency) without CSA treatment. Recipients were sexual mature and underwent bilateral ovariectomy before transplantation. The donor ovarian tissue were transplanted into subrenal capsule of the recipients. BLI was analyzed by the IVIS Imaging System 50, recipient mice were anesthetized, injected luciferin. Series in vivo imaging were assessed from day 1 to day 56 after transplantation in a quantified and time-course manner. Immunohistochemical staining of CD8+/CD4+-positive cells in ovarian graft and flow cytometry analysis of subpopulation T cells in the peripheral blood were assayed for rejection. Ovarian hormone activity was evaluated by vaginal cytology. RESULTS: The trend of BLI photon quantity declined significantly between the 1st day to the 5th day after transplantation. The steepest slope declined in group 4. It revealed mild increase trend between the 5th to 9th day but the trend declined again between the 9th day afterward in group 2, 3 and 4 but increased in group 1. Among the latter four groups in the second decline trend, the steepest slope occurred in group 4. Time by group effect showed the most significant declined trend of BLI signal in group 4 compared with the least change in group 1 (P=0.0007). Ovarian function and reserve remain constant in group 1, 2 and 5 after 56-day track. CONCLUSIONS: BLI may expedite the fast throughput screening of engraftment by a longitudinal monitoring in living animals to facilitate the study of the physiology and function of grafts, and the evaluation of new transplantation protocols in a rapid and reliable fashion. The pattern of engraftment of iso- and allo-ovarian graft existed differently depending on genetic background and the use of immunosuppressive therapy.