Organotypic stromal cells impact endothelial cell transcriptome in 3D microvessel networks
Matthew B Curtis1, Natalie Kelly1, Christopher C W Hughes2,3
1Department of Biomedical Engineering, University of California, Davis, 451 E. Health Sciences Drive, Room 2315, Davis, CA, 95616, USA.
Scientific Reports
|November 28, 2022
Summary
Stromal cells guide endothelial cell differentiation in 3D microvessel networks, creating organ-specific phenotypes. This organotypic model more accurately reflects in vivo endothelial cell behavior than traditional 2D cultures.
Area of Science:
- Vascular Biology
- Cell Biology
- Tissue Engineering
Background:
- Endothelial cells (ECs) exhibit organ-specific phenotypes despite performing similar functions.
- Mechanisms driving EC organ differentiation are not fully understood.
- Understanding EC organotropism is crucial for regenerative medicine and disease modeling.
Purpose of the Study:
- To investigate the role of stromal cells in driving organ-specific endothelial cell differentiation.
- To develop a 3D in vitro model that recapitulates in vivo microvessel characteristics.
- To compare the transcriptomic profiles of in vitro-generated ECs with their in vivo counterparts.
Main Methods:
- Generation of 3D microvessel networks using a common naïve endothelial cell and six different organ-derived stromal cells.
- Single-cell RNA sequencing (scRNA-Seq) to analyze EC populations and stromal cell subpopulations.
- Morphological analysis of organotypic vessel networks.
- Transcriptomic comparison between 3D in vitro models and in vivo ECs.
Main Results:
- scRNA-Seq identified five distinct EC populations, with proportions varying based on stromal cell type.
- Morphological network features inversely correlated with ECs involved in protein synthesis.
- Stromal cells exhibited unique subpopulations for extracellular matrix organization.
- 3D in vitro EC transcriptomes more closely mirrored in vivo ECs from corresponding organs (heart, skin, lung, pancreas).
Conclusions:
- Stromal cells significantly contribute to endothelial cell and microvessel network organ tropism.
- 3D organotypic models generate EC phenotypes that better resemble in vivo conditions.
- This study provides a novel platform for studying organ-specific vascular biology and developing targeted therapies.
