Long‐Term in Vitro Function of Adult Hepatocytes in a Collagen Sandwich Configuration

TLDR

Adult rat hepatocytes were sandwiched between two layers of hydrated rat tail tendon collagen matrix to reconstruct liver cellular polarity. In the sandwich system, hepatocytes retained secretion of albumin, transferrin, fibrinogen, bile acids, and urea for at least six weeks, whereas single-layer cultures lost these functions within 1–2 weeks; adding a second collagen layer after one week restored function, and the substrate type was irrelevant provided it supported attachment, with actin organization resembling in vivo and stress fibers absent, showing that extracellular configuration profoundly affects hepatocyte behavior.

Abstract

Abstract In an effort to reconstruct the cellular polarity normally found in the liver, adult rat hepatocytes were sandwiched between two layers of hydrated rat tail tendon collagen matrix. Functionally, sandwiched hepatocytes maintained the secretion of albumin, transferrin, fibrinogen, bile acids, and urea for at least 6 weeks, whereas cells cultured on a single layer of collagen gel ceased such secretion in 1–2 weeks. After 1 week of culture on a single layer of collagen gel, hepatocytes could still recover these lost functions when a second layer of collagen gel was applied. The exact nature of the substrate for constructing the sandwich system appeared to be unimportant as long as it allowed cellular attachment. Hepatocytes cultured in the sandwich system appeared to maintain a distribution of actin filaments similar to the in vivo state, whereas cells cultured on a single layer of collagen gel showed abnormal formation of stress fibers. These studies suggest that simple manipulations of the configuration of extracellular elements can dramatically alter the behavior of cultured hepatocytes.