Point-of-use, automated fabrication of a 3D human liver model supplemented with human adipose microvessels

Authors:
Sarah M Moss , Jillian Schilp, Maya Yaakov , Madison Cook , Erik Schuschke , Brandon Hanke , Hannah A Strobel , James B Hoying 
In:
Source: SLAS Discov
Publication Date: ()
Issue: S2472: 13680-4
Research Area:
Toxicology
Drug Discovery
Cells used in publication:
Hepatocyte, rat
Species: rat
Tissue Origin: liver
Experiment

Rat vascularized liver tissue construct
To demonstrate flexibility of the assay rodent hepatocytes (Lonza) and rodent NPCs (Lonza) with human adipose microvessels (haMVs, Advanced Solutions Life Sciences) were used. In the case of static cultures, the same fabrication strategy as described for human liver constructs was used with hepatocytes seeded at 10M cells/ml and NPCs at 1M cells/ml. Constructs were cultured for 1 week prior to the drug treatment. 

Abstract

Advanced in vitro tissue models better reflect healthy and disease tissue conditions in the body. However, complex tissue models are often manufactured using custom solutions and can be challenging to manufacture to scale. Here, we describe the automated fabrication of a cell-dense, thick (= 1 cm), human vascularized liver tissue model using a robotic biomanufacturing platform and off-the-shelf components to build, culture, and sample liver tissues hands-free without compromising tissue health or function. Fabrication of the tissue involved 3D bioprinting and incorporation of primary human hepatocytes, primary human non-parenchymal cells, and isolated fragments of intact human microvessels as vascular precursors. No differences were observed in select assessments of the liver tissues fabricated by hand or via automation. Furthermore, constant media exchange, via perfusion, improved urea output and elevated tissue metabolism. Interestingly, inclusion of adipose-derived human microvessels enhanced functional gene expression, including an enhanced response to a drug challenge. Our results describe the fabrication of a thick liver tissue environment useful for a variety of applications including liver disease modeling, infectious agent studies, and cancer investigations. We expect the automated fabrication of the vascularized liver tissue, at the point of use and using off-the-shelf platforms, eases fabrication of the complex model and increases its utility.