Decellularized Liver Nanofibers Enhance Liver Cell Using Non-Synthetic Polymer Blends

At a Glance

In vitro human liver models are utilized for screening pharmaceuticals and industrial chemicals, mimicking the key aspects of liver diseases for the discovery of novel molecular targets, and building tissue surrogates for implantation into patients suffering from end-stage liver failure. However, current models for growing and maintaining liver cells are not sufficient for long term usage. This work developed at Colorado State University has shown than liver cells can be cultured up to 47 days.

Background

Primary human hepatocytes (PHHs) are often used to fabricate models of liver disease given their ability to perform most liver functions such as protein synthesis, bile production, glucose and fatty acid metabolism, and the detoxification of endogenous and exogenous substances; however, PHH rapidly lose functions when cultured on their own on collagen I adsorbed onto non-physiological stiff substrates (e.g., glass and polystyrene). While the PHH phenotype can be stabilized in vitro for several weeks upon co-cultivation with certain non-parenchymal cell (NPC) types, functions still remain below those in freshly isolated PHHs typically due to the use of non-physiologic substrates. Thus, there remains a critical need to develop more physiologic substrates for PHH and PHH/NPC co-cultures alike.

Overview

This technology was developed to electrospin stable nanofibers of decellularized porcine liver ECM (PLECM) given their robust availability and the ability to support some hepatocyte functions in vitro; as control ECM, we utilized rat tail collagen-I, widely used for PHH culture due to ready availability and cost-effectiveness, and the polysaccharide, chitosan, that has been previously shown to stabilize collagen scaffolds and support some level of hepatic functions. We then cultured PHHs on their own and with different well-established NPC types, namely 3T3-J2 murine embryonic fibroblasts and primary human liver sinusoidal endothelial cells (LSECs), on the different nanofibrous scaffolds and non-fibrous control substrates; cell morphology, viability, and phenotypic functions were assessed for up to 47 days in culture.

Benefits

  • Long term use of primary liver cells to screen pharmaceutical drugs
  • Nanofiber morphology remains intact leading to suitable liver culture and phenotypic assessments
  • More in vivo like cultures

Applications

  • Screening of pharmaceutical drugs
  • No need for synthetic polymers
  • Application to human and other mammalian liver cells
Last Updated: September 2023
Opportunity

Available for Licensing
TRL: 5

IP Status

Provisional Patent Filed

Inventors

Matt Kipper
Liszt Coutinho Madruga
Jennifer Liu
Salman Khetani

Reference Number
2022-031
Licensing Manager

Steve Foster
Steve.Foster@colostate.edu
970-491-7100