Gutsche A T, Lo H, Zurlo J, Yager J, Leong K W
Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
Biomaterials. 1996 Feb;17(3):387-93. doi: 10.1016/0142-9612(96)85577-3.
Many tissue engineering applications require a scaffold or template conducive to cell attachment and maintenance of functions. It may also be advantageous in some cases for these scaffolds to have a controlled porous architecture to facilitate cellular or tissue ingrowth. In this study, we have engineered a porous carbohydrate-derivatized substrate for hepatocyte culture. Polystyrene foams, with pore sizes up to 100 microns, fabricated by phase separation from a homogeneous naphthalene solution, were derivatized with lactose and heparin, both of which are known to promote rat hepatocyte attachment and maintenance of its differentiated functions. Rat hepatocytes cultured on these derivatized foams exhibited a rounded cellular morphology with many microvilli evident on the surface of the cells. The hepatocytes showed an increase in albumin secretion for the first 3 days of culture in a defined, serum-free medium, and dropped back to initial levels by the end of 7 days. The production of cytochrome P450-dependent hydroxytestosterone metabolites were also measured. Two testosterone metabolites were maintained and five others were present but decreased over a culture period of 1 week. These carbohydrate-derivatized porous substrates may be useful for large-scale culture of hepatocytes, toxicology screening and for use in a liver assist device.
许多组织工程应用都需要一种有利于细胞附着和功能维持的支架或模板。在某些情况下，这些支架具有可控的多孔结构以促进细胞或组织向内生长可能也是有利的。在本研究中，我们设计了一种用于肝细胞培养的多孔碳水化合物衍生化基质。通过从均匀的萘溶液中相分离制备的孔径达100微米的聚苯乙烯泡沫，用乳糖和肝素进行衍生化，已知这两种物质都能促进大鼠肝细胞附着并维持其分化功能。在这些衍生化泡沫上培养的大鼠肝细胞呈现出圆形细胞形态，细胞表面可见许多微绒毛。在限定的无血清培养基中培养的前3天，肝细胞白蛋白分泌增加，到第7天结束时回落至初始水平。还测定了细胞色素P450依赖的羟基睾酮代谢物的产生。在1周的培养期内，两种睾酮代谢物得以维持，另外五种存在但减少。这些碳水化合物衍生化的多孔基质可能有助于肝细胞的大规模培养、毒理学筛选以及用于肝辅助装置。
