Rohn Susanne, Schroeder Jan, Riedel Henriette, Polenz Dietrich, Stanko Katarina, Reutzel-Selke Anja, Tang Peter, Brusendorf Lydia, Raschzok Nathanael, Neuhaus Peter, Pratschke Johann, Sawitzki Birgit, Sauer Igor M, Mogl Martina T
General, Visceral and Transplantation Surgery, Experimental Surgery and Regenerative Medicine, Campus Virchow-Klinikum, Berlin, Germany.
Cells Tissues Organs. 2015;201(6):399-411. doi: 10.1159/000445792. Epub 2016 May 20.
Stable long-term functioning of liver cells after transplantation in humans is still not achieved successfully. A new approach for successful engraftment of liver cells may be the transplantation of syngeneic cells into an allogeneic liver graft. We therefore developed a new rat model for combined liver and liver cell transplantation (cLCTx) under stable immunosuppression.
After inducing a mitotic block, liver grafts from female donor rats (Dark Agouti) were transplanted into female recipients (Lewis). In male Lewis rats, liver cell proliferation was induced with subsequent cell isolation and transplantation into female recipients after organ transplantation. Y-chromosome detection of the transplanted male cells was performed by quantitative polymerase chain reaction (qPCR) and fluorescence in situ hybridization (FisH) with localization of transplanted cells by immunohistochemistry.
Immunohistochemistry demonstrated the engraftment of transplanted cells, as confirmed by FisH, showing repopulation of the liver graft with 15.6% male cells (± 1.8 SEM) at day 90. qPCR revealed 14.15% (± 5.09 SEM) male DNA at day 90.
Engraftment of transplanted syngeneic cells after cLCTx was achieved for up to 90 days under immunosuppression. Immunohistochemistry indicated cell proliferation, and the FisH results were partly confirmed by qPCR. This new protocol in rats appears feasible for addressing long-term functioning and eventually the induction of operational tolerance in the future.
人类肝脏细胞移植后仍未成功实现长期稳定功能。肝细胞成功植入的一种新方法可能是将同基因细胞移植到同种异体肝脏移植物中。因此，我们开发了一种在稳定免疫抑制下进行联合肝脏和肝细胞移植（cLCTx）的新大鼠模型。
诱导有丝分裂阻滞之后，将雌性供体大鼠（深色刺鼠）的肝脏移植物移植到雌性受体（刘易斯大鼠）体内。在雄性刘易斯大鼠中，诱导肝细胞增殖，随后进行细胞分离，并在器官移植后将细胞移植到雌性受体中。通过定量聚合酶链反应（qPCR）和荧光原位杂交（FisH）对移植的雄性细胞进行Y染色体检测，并通过免疫组织化学对移植细胞进行定位。
免疫组织化学证实了移植细胞的植入，FisH也证实了这一点，显示在第90天时肝脏移植物中有15.6%的雄性细胞（±1.8标准误）重新填充。qPCR显示在第90天时雄性DNA为14.15%（±5.·09标准误）。
在免疫抑制下，cLCTx后移植的同基因细胞可成功植入长达90天。免疫组织化学表明细胞增殖，FisH结果部分得到qPCR的证实。这种新的大鼠实验方案似乎对于解决长期功能以及最终在未来诱导手术耐受是可行的。
