Departments of Chemical Engineering and Materials Science, University of Minnesota, 55455-0132, Minneapolis, MN, USA.
Cytotechnology. 2004 Jul;45(3):125-40. doi: 10.1007/s10616-004-7996-6.
Hepatocytes self-assemble in culture to form compacted spherical aggregates, or spheroids, that mimic the structure of the liver by forming tight junctions and bile canalicular channels. Hepatocyte spheroids thus resemble the liver to a great extent. However, liver tissue contains other cell types and has bile ducts and sinusoids formed by endothelial cells. Reproducing 3-D co-culture in vitro could provide a means to develop a more complex tissue-like structure. Stellate cells participate in revascularization after liver injury by excreting between hepatocytes a laminin trail that endothelial cells follow to form sinusoids. In this study we investigated co-culture of rat hepatocytes and a rat hepatic stellate cell line, HSC-T6. HSC-T6, which does not grow in serum-free spheroid medium, was able to grow under co-culture conditions. Using a three-dimensional cell tracking technique, the interactions of HSC-T6 and hepatocyte spheroids were visualized. The two cell types formed heterospheroids in culture, and HSC-T6 cell invasion into hepatocyte spheroids and subsequent retraction was observed. RT-PCR revealed that albumin and cytochrome P450 2B1/2 expression were better maintained in co-culture conditions. These three-dimensional heterospheroids provide an attractive system for in vitro studies of hepatocyte-stellate cell interactions.
肝细胞在培养中自组装形成紧密的球形聚集体，即球体，通过形成紧密连接和胆小管通道，模拟肝脏的结构。因此，肝细胞球体在很大程度上类似于肝脏。然而，肝组织还包含其他细胞类型，并且具有由内皮细胞形成的胆管和窦隙。在体外复制 3D 共培养可能提供一种方法来开发更复杂的组织样结构。肝星状细胞在肝损伤后通过在肝细胞之间分泌层粘连蛋白轨迹参与再血管化，内皮细胞沿着该轨迹形成窦隙。在这项研究中，我们研究了大鼠肝细胞和大鼠肝星状细胞系 HSC-T6 的共培养。HSC-T6 在无血清球体培养基中不能生长，但在共培养条件下能够生长。使用三维细胞跟踪技术，可视化了 HSC-T6 和肝细胞球体的相互作用。两种细胞类型在培养中形成异质球体，并且观察到 HSC-T6 细胞侵入肝细胞球体和随后的回缩。RT-PCR 显示，在共培养条件下白蛋白和细胞色素 P450 2B1/2 的表达得到更好的维持。这些三维异质球体为体外研究肝细胞-星状细胞相互作用提供了一个有吸引力的系统。
