TY - JOUR
T1 - Optimising a self-assembling peptide hydrogel as a Matrigel alternative for 3-dimensional mammary epithelial cell culture
AU - Lingard, Eliana
AU - Dong, Siyuan
AU - Hoyle, Anna
AU - Appleton, Ellen
AU - Hales, Alis
AU - Skaria, Eldhose
AU - Lawless, Craig
AU - Taylor-Hearn, Isobel
AU - Saadati, Simon
AU - Chu, Qixun
AU - Miller, Aline F
AU - Domingos, Marco
AU - Saiani, Alberto
AU - Swift, Joe
AU - Gilmore, Andrew P
N1 - Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.
PY - 2024/6/1
Y1 - 2024/6/1
N2 - Three-dimensional (3D) organoid models have been instrumental in understanding molecular mechanisms responsible for many cellular processes and diseases. However, established organic biomaterial scaffolds used for 3D hydrogel cultures, such as Matrigel, are biochemically complex and display significant batch variability, limiting reproducibility in experiments. Recently, there has been significant progress in the development of synthetic hydrogels for in vitro cell culture that are reproducible, mechanically tuneable, and biocompatible. Self-assembling peptide hydrogels (SAPHs) are synthetic biomaterials that can be engineered to be compatible with 3D cell culture. Here we investigate the ability of PeptiGel® SAPHs to model the mammary epithelial cell (MEC) microenvironment in vitro. The positively charged PeptiGel®Alpha4 supported MEC viability, but did not promote formation of polarised acini. Modifying the stiffness of PeptiGel® Alpha4 stimulated changes in MEC viability and changes in protein expression associated with altered MEC function, but did not fully recapitulate the morphologies of MECs grown in Matrigel. To supply the appropriate biochemical signals for MEC organoids, we supplemented PeptiGels® with laminin. Laminin was found to require negatively charged PeptiGel® Alpha7 for functionality, but was then able to provide appropriate signals for correct MEC polarisation and expression of characteristic proteins. Thus, optimisation of SAPH composition and mechanics allows tuning to support tissue-specific organoids.
AB - Three-dimensional (3D) organoid models have been instrumental in understanding molecular mechanisms responsible for many cellular processes and diseases. However, established organic biomaterial scaffolds used for 3D hydrogel cultures, such as Matrigel, are biochemically complex and display significant batch variability, limiting reproducibility in experiments. Recently, there has been significant progress in the development of synthetic hydrogels for in vitro cell culture that are reproducible, mechanically tuneable, and biocompatible. Self-assembling peptide hydrogels (SAPHs) are synthetic biomaterials that can be engineered to be compatible with 3D cell culture. Here we investigate the ability of PeptiGel® SAPHs to model the mammary epithelial cell (MEC) microenvironment in vitro. The positively charged PeptiGel®Alpha4 supported MEC viability, but did not promote formation of polarised acini. Modifying the stiffness of PeptiGel® Alpha4 stimulated changes in MEC viability and changes in protein expression associated with altered MEC function, but did not fully recapitulate the morphologies of MECs grown in Matrigel. To supply the appropriate biochemical signals for MEC organoids, we supplemented PeptiGels® with laminin. Laminin was found to require negatively charged PeptiGel® Alpha7 for functionality, but was then able to provide appropriate signals for correct MEC polarisation and expression of characteristic proteins. Thus, optimisation of SAPH composition and mechanics allows tuning to support tissue-specific organoids.
KW - Laminin/pharmacology
KW - Hydrogels/chemistry
KW - Drug Combinations
KW - Proteoglycans/pharmacology
KW - Collagen/chemistry
KW - Peptides/pharmacology
KW - Epithelial Cells/drug effects
KW - Humans
KW - Female
KW - Cell Culture Techniques, Three Dimensional/methods
KW - Cell Survival/drug effects
KW - Biocompatible Materials/chemistry
KW - Mammary Glands, Human/cytology
KW - Organoids/drug effects
KW - Cell Culture Techniques/methods
UR - https://pubmed.ncbi.nlm.nih.gov/38657288/
U2 - 10.1016/j.bioadv.2024.213847
DO - 10.1016/j.bioadv.2024.213847
M3 - Article
C2 - 38657288
SN - 2772-9508
VL - 160
JO - Biomaterials Advances
JF - Biomaterials Advances
M1 - 213847
ER -