Peptide Hydrogels for 3d Hepatocyte Encapsulation and Function

The relationship of self-assembling peptide hydrogel (SAPHs) material properties with hepatocyte viability, metabolism and morphology using a series of PeptiGel® SAPHs is presented. Negatively charged SAPHs promote higher cell viability and proliferation for hepatocytes rather than positively charged SAPHs. Alpha7, a novel negatively charged hydrogel constitutes an ideal environment in vitro for the 3D culture of hepatocytes, supporting major hepatic functions and differentiated morphology over 14 days. Specifically, 3D cultured HepG2 cells within Alpha7, lead to a significant enhancement of albumin protein production (4.4-fold) and albumin mRNA level, as well as maintenance of CYP450s enzyme activities (CYP1A2, CYP2C9, CYP2E1, CYP3A4) over 14 days. 3D hepatocytes morphology and ECM protein deposition are investigated. Fluorescent/ICC staining (DAPI/F-actin/E-cadherin) showed that HepG2 cell spheroids within Alpha7 maintain a differentiated phenotype and form stronger cell-cell interactions over 14 days. Furthermore, 3D SAPH cultured HepG2 cells show response to 2mM acetaminophen (CYP2E1 gene highly induced). These findings provide new information about the use of negatively charged SAPHs in 3D culture for hepatocytes, with potential applications in drug screening and liver tissue regeneration. © 2023, The Authors. All rights reserved.