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The In Vivo Imaging Platform for Cell Tracking in Vascularized Composite Allotransplantation
Chih-jen Wen, PhD; H.Y. Cheng, PhD; Y.L. Wang, PhD; Ling-Yi Shih; Chih-Fan Lin; Xiao-Ting Huang; FC Wei
Chang Gung Memorial Hospital, Taoyuan, Taiwan
Structural/functional recovery and overcoming rejection are two main obstacles of vascularized composite allotransplantation (VCA; also known as "composite tissue allotransplant"). Therapies with the Schwann's cells, adipocyte-derived stem cells (ADSC), and regulatory T cells (Treg) were used for structural/functional recovery and overcoming rejection. These cells for therapies and cells related to structural/functional recovery and rejection were tracked by in vivo optical imaging with ex vivo and in vitro validation. "Cell tracking" means that in vivo tracking the migration, homing, behavior and fate of the specific live cell(s) by imaging techniques. Long-term observation is indispensable for the research of recovery and rejection with their therapies. The traditional observation methods, biopsy and suspended single cells (e.g. blood samples or isolated cells from tissues by enzymatic digestion), are flawed with sampling bias and a lack of dynamic real time inspection; and more importantly, they are both invasive. Biopsy and blood sampling methods are destructive detections because that the immunity, recovery or cell movement might be affected by the wound and the correctness of the research might be affected consequently. On this account, the development of an in vivo imaging platform for long-term monitoring is urgently needed. Nonetheless, the short follow-up time provided by the mainstream live animal imaging techniques has restricted their application for in vivo cell tracking purposes. Optical imaging, on the other hand, provides long-term tracking, high sensitivity and real-time imaging. In this study, the in vivo dynamic biodistribution of cells for therapies and the cells related to recovery/rejection were optical imaged with bioluminescence from the Luciferase transgenic rat and fluorescence from the near infrared fluorescence probe. To confirm the effectiveness and accuracy of this platform, in vitro validation was completed with objective, accurate and reproducible "tissomic" cellular quantitative analysis from the new technology "Tissomics Analyzer (TissueGnostics TissueFAXS PlusTM)". After being in vivo imaged, the VCA rat was whole-body sectioned for tissomic validation and mechanism studies. For example, as shown in the attached figure, the major targets of the ADSC are lung, liver, spleen and the transplanted allograft. The in vivo imaging platform is not only important for the research of mechanisms, but also facilitates the screening of therapies for overcoming rejection; it will provide valuable information for both academic research and clinical application.
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