Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Shinjuku-ku, Tokyo, Japan.
Cell Transplant. 2012;21(2-3):429-36. doi: 10.3727/096368911X605330.
Recent advances in tissue engineering technologies have highlighted the ability to create functional liver systems using isolated hepatocytes in vivo. Considering the serious shortage of donor livers that can be used for hepatocyte isolation, it has remained imperative to establish a hepatocyte propagation protocol to provide highly efficient cell recovery allowing for subsequent tissue engineering procedures. Donor primary hepatocytes were isolated from human α-1 antitrypsin (hA1AT) transgenic mice and were transplanted into the recipient liver of urokinase-type plasminogen activator-severe combined immunodeficiency (uPA/SCID) mice. Transplanted donor hepatocytes actively proliferated within the recipient liver of the uPA/SCID mice. At week 8 or later, full repopulation of the uPA/SCID livers with the transplanted hA1AT hepatocytes were confirmed by blood examination and histological assessment. Proliferated hA1AT hepatocytes were recovered from the recipient uPA/SCID mice, and we generated hepatocyte sheets using these recovered hepatocytes for subsequent transplantation into the subcutaneous space of mice. Stable persistency of the subcutaneously engineered liver tissues was confirmed for up to 90 days, which was the length of our present study. These new data demonstrate the feasibility in propagating murine hepatocytes prior to the development of hepatic cells and bioengineered liver systems. The ability to regenerate and expand hepatocytes has potential clinical value whereby procurement of small amounts of tissue could be expanded to sufficient quantities prior to their use in hepatocyte transplantation or other hepatocyte-based therapies.
最近组织工程技术的进展突出了使用体内分离的肝细胞创建功能性肝脏系统的能力。考虑到可用于分离肝细胞的供体肝脏严重短缺，建立肝细胞增殖方案以提供高效的细胞回收，从而允许进行后续的组织工程程序仍然至关重要。从人α-1 抗胰蛋白酶（hA1AT）转基因小鼠中分离供体原代肝细胞，并将其移植到尿激酶型纤溶酶原激活物-严重联合免疫缺陷（uPA/SCID）小鼠的受体肝脏中。移植的供体肝细胞在 uPA/SCID 小鼠的受体肝脏中积极增殖。在第 8 周或之后，通过血液检查和组织学评估确认 uPA/SCID 肝脏中完全用移植的 hA1AT 肝细胞重新填充。从受体 uPA/SCID 小鼠中回收增殖的 hA1AT 肝细胞，并使用这些回收的肝细胞生成肝细胞片，用于随后移植到小鼠的皮下空间。在我们目前的研究中，证实了皮下工程化肝组织长达 90 天的稳定持续存在。这些新数据表明，在开发肝细胞和生物工程肝脏系统之前，繁殖小鼠肝细胞是可行的。再生和扩大肝细胞的能力具有潜在的临床价值，因为可以在将其用于肝细胞移植或其他基于肝细胞的治疗之前，从小量组织中扩增到足够的数量。
