Long-term maintenance of functional primary human hepatocytes in 3D gelatin matrices produced by solution blow spinning

Abstract

Solution blow spinning (SBS) has recently emerged as a novel method that can produce nano- and microfiber structures suitable for tissue engineering. Gelatin is an excellent precursor for SBS as it is derived mainly from collagens that are abundant in natural extracellular matrices. Here we report, for the first time the successful generation of 3D thermally crosslinked preforms by using SBS from porcine gelatin. These SBS mats were shown to have three-dimensional fibrous porous structure similar to that of mammalian tissue extracellular matrix. In pharma industry, there is an urgent need for adequate 3D liver tissue models that could be used in high throughput setting for drug screening and to assess drug induced liver injury. We used SBS mats as culturing substrates for human hepatocytes to create an array of 3D human liver tissue equivalents in 96-well format. The SBS mats were highly cytocompatible, facilitated the induction of hepatocyte specific CYP gene expression in response to common medications, and supported the maintenance of hepatocyte differentiation and polarization status in long term cultures for more than 3 weeks. Together, our results show that SBS-generated gelatin scaffolds are a simple and efficient platform for use in vitro for drug testing applications.

Fabrication of oxygen‐carrying microparticles functionalized with liver ECM‐proteins to improve phenotypic three‐dimensional in vitro liver assembly, function, and responses

“…We did not find any significant differences (p > 0.05) between laminin-111 conjugated and noncoated MPs in terms of liver-specific functions probed, highlighting the importance of proteins native to the mature in vivo liver environment for benefitting cell culture in vitro. Our result is consistent with an experiment conducted by Klaas et. al (2021) where they cultured HepG2 cells on 3D nano-and microfiber structures based on gelatin, preincubated with laminin-111 and observed no significant changes in cell morphology, viability, and production of proteins between coated with laminin-111 and uncoated 3D gelatin scaffolds.…”

“…Our result is consistent with an experiment conducted by Klaas et. al (2021) where they cultured HepG2 cells on 3D nano‐ and microfiber structures based on gelatin, preincubated with laminin‐111 and observed no significant changes in cell morphology, viability, and production of proteins between coated with laminin‐111 and uncoated 3D gelatin scaffolds.…”

Fabrication of oxygen carrying microparticles functionalized with liver ECM-proteins to improve phenotypic three-dimensional in vitro liver assembly, function, and responses

“…Our result is consistent with an experiment conducted by Klaas et. al [46] where they cultured HepG2 cells on 3D nano-and microfiber structures based on gelatin, preincubated with laminin-111 and observed no significant changes in cell morphology, viability and production of proteins between coated with laminin-111 and uncoated 3D gelatin scaffolds.…”

The native liver as inspiration to create superior in vitro hepatic models

“…207 A very important feature of in vitro DILI models involves their long-term cell viability and functionality, which is only preserved during several hours in case of conventional 2D-cultures resulting in a limited presence of phase I and II metabolic enzymes. 37 Within this short term, it is not possible to track the toxicity of drug compounds that lead to enzyme induction, immune-mediated toxicity (i.e. idiosyncratic DILI -iDILI) or compounds with a low clearance rate.…”