Demetriou A A, Whiting J, Levenson S M, Chowdhury N R, Schechner R, Michalski S, Feldman D, Chowdhury J R
Ann Surg. 1986 Sep;204(3):259-71. doi: 10.1097/00000658-198609000-00005.
A technique has been developed by the authors that allows hepatocyte attachment on collagen-coated microcarriers resulting in prolonged hepatocyte viability and function both in vivo and in vitro. Rat hepatocytes were obtained by portal vein collagenase perfusion. Intraperitoneally transplanted microcarrier-attached normal hepatocytes into congeneic Gunn rats were functioning 3-4 weeks later, as shown by the presence and persistence of conjugated bilirubin in recipient bile, sustained decrease in serum bilirubin, uptake of Tc99m-DESIDA, and morphologic criteria. Intraperitoneal transplantation of normal microcarrier-attached hepatocytes into genetically albumin deficient rats (NAR) resulted in marked increase in plasma albumin levels (6 days without and 21 days with Cyclosporin A immunosuppression). Microcarrier-attached hepatocytes transplanted after 2 weeks of storage at -80 C into congeneic Gunn rats were viable and functional as assessed by criteria outlined above. An extracorporeal liver perfusion system was developed using the microcarrier-attached hepatocytes that was capable of synthesizing and conjugating bilirubin and synthesizing liver-specific proteins.
作者开发了一种技术，可使肝细胞附着于胶原包被的微载体上，从而在体内和体外均能延长肝细胞的存活时间并维持其功能。通过门静脉胶原酶灌注获取大鼠肝细胞。将微载体附着的正常肝细胞经腹腔移植到同基因的Gunn大鼠体内，3 - 4周后这些肝细胞开始发挥功能，表现为受体胆汁中结合胆红素的出现和持续存在、血清胆红素持续下降、Tc99m - DESIDA摄取以及形态学标准。将微载体附着的正常肝细胞经腹腔移植到遗传性白蛋白缺乏大鼠（NAR）体内，血浆白蛋白水平显著升高（未使用环孢素A免疫抑制时为6天，使用环孢素A免疫抑制时为21天）。在 - 80℃储存2周后的微载体附着肝细胞经移植到同基因的Gunn大鼠体内，根据上述标准评估仍具有活力且功能正常。利用微载体附着肝细胞开发了一种体外肝脏灌注系统，该系统能够合成和结合胆红素并合成肝脏特异性蛋白质。
