A Whole-Organ Regenerative Medicine Approach for Liver Replacement
A Whole-Organ Regenerative Medicine Approach for Liver Replacement
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DOI：
10.1089/ten.tec.2010.0698
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发表时间：
2011-06-01
影响因子：
3
通讯作者：
Badylak, Stephen F.
中科院分区：
文献类型：
--
作者：
Soto-Gutierrez, Alejandro;Zhang, Li;Badylak, Stephen F.
背景和目标：终末期肝病的治疗选择是全器官肝移植，但这种选择受到供体器官短缺的限制。基于细胞的治疗和肝组织工程已被认为是肝移植的替代方案，但迄今为止都没有被证明有效。最近已经描述了用于肝脏替代的再生医学方法，其包括使用通过异种肝脏的去细胞化制备的三维器官支架。本研究探讨了一种新的，最小的破坏性的方法，全器官肝脱细胞和三种不同的细胞重新接种策略，以工程师的功能liver tissue.Methods：酶，洗涤剂和机械方法的组合被用来删除所有的细胞从离体大鼠肝脏。在定制的器官室中使用全器官灌注，并通过形态学、生物化学和免疫标记技术检查脱细胞化的肝脏，以保存天然基质结构和组成。评价了三种不同的用于所得三维肝脏支架的肝细胞接种的方法以最大化细胞存活和功能：（1）直接实质注射，（2）多步输注，或（3）连续灌注。去细胞化过程保留了三维宏观结构、超微结构、细胞外基质组分的组成、肝脏的天然微血管网络，和胆汁引流系统，以及高达50%的生长因子含量。三维肝基质resstrike与肝细胞的多步输注产生类似于90%的细胞植入和支持肝脏特定的功能能力的移植细胞，包括白蛋白的生产，尿素代谢，和细胞色素P450 induction.Conclusions：全器官肝脱细胞是可能的，与维护的结构和组成适合支持功能肝细胞。
Background & Aims: The therapy of choice for end-stage liver disease is whole-organ liver transplantation, but this option is limited by a shortage of donor organs. Cell-based therapies and hepatic tissue engineering have been considered as alternatives to liver transplantation, but neither has proven effective to date. A regenerative medicine approach for liver replacement has recently been described that includes the use of a three-dimensional organ scaffold prepared by decellularization of xenogeneic liver. The present study investigates a new, minimally disruptive method for whole-organ liver decellularization and three different cell reseeding strategies to engineer functional liver tissue.Methods: A combination of enzymatic, detergent, and mechanical methods are used to remove all cells from isolated rat livers. Whole-organ perfusion is used in a customized organ chamber and the decellularized livers are examined by morphologic, biochemical, and immunolabeling techniques for preservation of the native matrix architecture and composition. Three different methods for hepatocyte seeding of the resultant three-dimensional liver scaffolds are evaluated to maximize cell survival and function: (1) direct parenchymal injection, (2) multistep infusion, or (3) continuous perfusion.Results: The decellularization process preserves the three-dimensional macrostructure, the ultrastructure, the composition of the extracellular matrix components, the native microvascular network of the liver, and the bile drainage system, and up to 50% of growth factor content. The three-dimensional liver matrix reseeded with the multistep infusion of hepatocytes generated similar to 90% of cell engraftment and supported liver-specific functional capacities of the engrafted cells, including albumin production, urea metabolism, and cytochrome P450 induction.Conclusions: Whole-organ liver decellularization is possible with maintenance of structure and composition suitable to support functional hepatocytes.