Chen Yibin, Geerts Sharon, Jaramillo Maria, Uygun Basak E
Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, 51 Blossom Street, Boston, MA, 02114, USA.
Shriners Hospitals for Children in Boston, 51 Blossom Street, Boston, MA, 02114, USA.
Methods Mol Biol. 2018;1577:255-270. doi: 10.1007/7651_2017_56.
Severe liver disease is the 12th leading cause of death in the USA, with organ transplantation often being the only viable option for treatment. However, due to the shortage of viable donor livers, it is estimated that over 1200 patients died in 2015 while waiting for liver transplantation. This highlights the need for alternative sources of viable organs. In this study, we describe a method that provides the groundwork for the development of functional liver grafts. The approach described here is for removal of cells from intact livers and subsequently repopulating them with functional liver cells. Briefly, rat livers are harvested and subjected to a series of perfusion decellularization steps using an anionic detergent such that an intact decellularized liver matrix (DLM) scaffold with preserved vascular architecture is obtained. Further, we describe methods to recellularize DLM scaffolds with adult primary hepatocytes, creating a liver graft that exhibits hepatic functions in vitro.
严重肝病是美国第12大死因，器官移植通常是唯一可行的治疗选择。然而，由于可用供体肝脏短缺，据估计2015年有超过1200名患者在等待肝移植时死亡。这凸显了寻找可用器官替代来源的必要性。在本研究中，我们描述了一种为功能性肝移植物的开发奠定基础的方法。这里描述的方法是从完整肝脏中去除细胞，随后用功能性肝细胞重新填充它们。简而言之，收获大鼠肝脏，并使用阴离子去污剂进行一系列灌注去细胞步骤，从而获得具有保留血管结构的完整去细胞肝脏基质（DLM）支架。此外，我们描述了用成年原代肝细胞使DLM支架重新细胞化的方法，从而创建一种在体外具有肝功能的肝移植物。
