Recent years have seen a proliferation of methods leading to successful organ decellularization. In this experiment we examine the feasibility of a decellularized liver construct to support growth of functional multilineage cells. Bio-chamber systems were used to perfuse adult rat livers with 0.1% SDS for 24 hours yielding decellularized liver scaffolds. Initially, we recellularized liver scaffolds using a human tumor cell line (HepG2, introduced via the bile duct). Subsequent studies were performed using either human tumor cells co-cultured with human umbilical vein endothelial cells (HUVECs, introduced via the portal vein) or rat neonatal cell slurry (introduced via the bile duct). Bio-chambers were used to circulate oxygenated growth medium via the portal vein at 37C for 5-7 days. Human HepG2 cells grew readily on the scaffold (n = 20). HepG2 cells co-cultured with HUVECs demonstrated viable human endothelial lining with concurrent hepatocyte growth (n = 10). In the series of neonatal cell slurry infusion (n = 10), distinct foci of neonatal hepatocytes were observed to repopulate the parenchyma of the scaffold. The presence of cholangiocytes was verified by CK-7 positivity. Quantitative albumin measurement from the grafts showed increasing albumin levels after seven days of perfusion. Graft albumin production was higher than that observed in traditional cell culture. This data shows that rat liver scaffolds support human cell ingrowth. The scaffold likewise supported the engraftment and survival of neonatal rat liver cell slurry. Recellularization of liver scaffolds thus presents a promising model for functional liver engineering.

近年来，导致成功的器官脱细胞的方法激增。在本实验中，我们研究了脱细胞肝构建体支持功能性多谱系细胞生长的可行性。生物腔室系统用于用0.1％SDS灌注成年大鼠肝脏24小时，产生脱细胞的肝支架。最初，我们使用人类肿瘤细胞系（HepG2，通过胆管引入）对肝支架进行了细胞重组。随后的研究是使用与人脐静脉内皮细胞（HUVEC，经门静脉引入）共培养的人肿瘤细胞或大鼠新生细胞浆液（经胆管引入）进行的。生物室用于在37℃下通过门静脉循环含氧的生长培养基5-7天。人HepG2细胞可在支架上轻松生长（n = 20）。与HUVECs共培养的HepG2细胞显示了可行的人类内皮细胞内膜，并发肝细胞生长（n = 10）。在一系列新生儿细胞浆输注（n = 10）中，观察到新生儿肝细胞的不同病灶重新填充了支架的实质。通过CK-7阳性证实了胆管细胞的存在。移植物中白蛋白的定量测量显示，灌注7天后白蛋白水平增加。嫁接白蛋白产量高于传统细胞培养中观察到的产量。该数据表明大鼠肝支架支持人类细胞向内生长。支架同样支持新生大鼠肝细胞浆液的植入和存活。肝支架的重新细胞化因此提出了功能性肝工程的有希望的模型。