Description

WP5 Cellular liver assays

Maintaining liver-specific functionality and phenotypicstability in vitro requires to (i) conserve the polarized structure, (ii) preserve cell-cell contacts and (iii) implement liver-specific non-parenchymal cells. Sandwich cultures represent the current gold standard where hepatocytes are grown in between layers of extracellular matrix. However, these cultures allow only for acute toxicity testing and isolation of RNA and protein isolation from these cultures has proven difficult. Reaggregation of hepatocytes to hepatospheres devoid of added scaffold materials, has shown to improve liver-specific functionality and in vitro live time to assess chronic toxicological effects. In addition, these re-aggregated liver tissue models are compatible with most of the assays envisioned within HeCaToS such as RNA and protein isolation and ROS measurement.

WP5 Cellular heart assays

Pluripotent stem cells have been shown in vitro to efficiently differentiate into spontaneously contracting cardiomyocyte-like cells34. Pluripotent stemcells have self-renewal capability in vitro and the capability to differentiate into all threegerm layers and thus, in principle, can give rise to all cell types of the human body. Human iPS cells, to be exploited by HeCaTos, have thus shown to be a valuable source for cardiomyocytes to establish in vitro assays.