Nanometric self-assembling peptide layers maintain adult hepatocyte phenotype in sandwich cultures

Background: Isolated hepatocytes removed from their microenvironment soon lose their hepatospecific functions when cultured. Normally hepatocytes are commonly maintained under limited culture medium supply as well as scaffold thickness. Thus, the cells are forced into metabolic stress that degenerate liver specific functions. This study aims to improve hepatospecific activity by creating a platform based on classical collagen sandwich cultures. Results: The modified sandwich cultures replace collagen with self-assembling peptide, RAD16-I, combined with functional peptide motifs such as the integrin-binding sequence RGD and the laminin receptor binding sequence YIG to create a cell-instructive scaffold. In this work, we show that a plasma-deposited coating can be used to obtain a peptide layer thickness in the nanometric range, which in combination with the incorporation of functional peptide motifs have a positive effect on the expression of adult hepatocyte markers including albumin, CYP3A2 and HNF4-alpha. Conclusions: This study demonstrates the capacity of sandwich cultures with modified instructive self-assembling peptides to promote cell-matrix interaction and the importance of thinner scaffold layers to overcome mass transfer problems. We believe that this bioengineered platform improves the existing hepatocyte culture methods to be used for predictive toxicology and eventually for hepatic assist technologies and future artificial organs. Background The liver is an important and complex organ that plays a vital role in metabolism and is responsible for many important functions of the body including glycogen storage, plasma protein production, drug detoxification and xenobiotics metabolization. Due to the importance of this organ in many of the body’s daily processes, liver malfunction often leads to death. Most of the activity of the liver can be attributed to hepatocytes, which make up 60-80% of the cytoplasmic mass of the liver [1,2]. Loss of hepatocyte function can result in acute or chronic liver disease and, as a result, substantially compromise the rest of the organ and the body. Many previous strategies have been implemented to maintain