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3D Culture of human lung fibroblasts decreases proliferative and increases extracellular matrix remodeling genes

ABSTRACT: 3D Culture of human lung fibroblasts decreases proliferative and increases extracellular matrix remodeling genes

PROVIDER: PRJNA1036649 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26713629_1.fastq.gz	Fastqsanger.gz
	SRR26713629_2.fastq.gz	Fastqsanger.gz
	SRR26713630_1.fastq.gz	Fastqsanger.gz
	SRR26713630_2.fastq.gz	Fastqsanger.gz
	SRR26713631_1.fastq.gz	Fastqsanger.gz

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3D Culture of human lung fibroblasts decreases proliferative and increases extracellular matrix remodeling genes

Project description:Fibroblasts are the main producers of extracellular matrix (ECM) responsible for ECM maintenance and repair, a process often disrupted in chronic lung diseases. The accompanying mechanical changes adversely affect resident cells and overall lung function. Numerous models have been used to elucidate fibroblast behavior which are now evolving towards complex 3D models incorporating ECM, aiming to replicate the cells’ native environment. Little is known about the cellular changes that occur when moving from 2D to 3D cell culture. This study compared gene expression profiles of primary human lung fibroblasts from 7 subjects with normal lung function, cultured for 24 hours on 2D collagen I-coated tissue culture plastic and in 3D collagen I hydrogels, which are commonly used to mimic ECM in various models, from contraction assays to organ-on-a-chip models. Comparing 3D to 2D cell culture, 6,771 differentially expressed genes (2,896 up, 3,875 down) were found; enriched gene sets within the downregulated genes, identified through Gene Set Enrichment Analysis and Ingenuity pathway analysis, were involved in the initiation of DNA replication which implied downregulation of fibroblast proliferation in 3D. Observation of cells for 72 hours in 2D and 3D environments confirmed the reduced progression through the cell cycle in 3D. A focused analysis examining the Hippo pathway and ECM-associated genes, showed differential patterns of gene expression in the 3D versus 2D culture. Altogether, the transcriptional response of fibroblasts cultured in 3D indicated inhibition of proliferation, and alterations in Hippo and ECM pathways indicating a complete switch from proliferation to ECM remodeling.

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