Summary
Published in International Journal of Molecular Sciences (2021), this peer-reviewed study used 3D Petri Dish® micro-molds to form scaffold-free 3D microtissues. Full citation: Messner, Catherine Jane, et al. Single Cell Gene Expression Analysis in a 3D Microtissue Liver Model Reveals Cell Type-Specific Responses to Pro-Fibrotic TGF-β1 Stimulation
Single Cell Gene Expression Analysis in a 3D Microtissue Liver Model Reveals Cell Type-Specific Responses to Pro-Fibrotic TGF-β1 Stimulation
Research Overview
Multicellular liver microtissues combining HepaRG, hTERT-HSC, and THP-1 cells maintain the interactions and physiological properties needed to model liver fibrosis — but that very complexity makes it hard to tell which cell type is responding.
This study applied single-cell RNA sequencing to separate the molecular responses of each cell type during fibrosis elicited by TGF-β1. An enzymatic dissociation method was optimized to obtain single-cell suspensions from the microtissues; the isolated cells retained good viability, could be re-plated and cultured in 2D, and expressed their expected profiles — validating the workflow.
Key Discoveries
- Single-cell RNA-seq resolved cell-type-specific responses within multicellular liver microtissues
- Optimized enzymatic dissociation preserved viability and re-plating capacity
- Applied to TGF-β1-induced fibrosis in a HepaRG/stellate/macrophage tri-culture model
