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Influence of three-dimensional culture in an interwoven hollow fiber bioreactor on the gene expression profile of hepatic HepG2 cells

ABSTRACT: Influence of three-dimensional culture in an interwoven hollow fiber bioreactor on the gene expression profile of hepatic HepG2 cells

PROVIDER: PRJNA545713 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR9166092_1.fastq.gz	Fastqsanger.gz
	SRR9166092_2.fastq.gz	Fastqsanger.gz
	SRR9166093_1.fastq.gz	Fastqsanger.gz
	SRR9166093_2.fastq.gz	Fastqsanger.gz
	SRR9166094_1.fastq.gz	Fastqsanger.gz

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Influence of three-dimensional culture in an interwoven hollow fiber bioreactor on the gene expression profile of hepatic HepG2 cells

Project description:Purpose: In this study, we aimed to elucidate the specific molecular mechanisms that are activated when hepatic cells are cultured in the 3D environment of an interwoven hollow fiber bioreactor device. Methods: HepG2 human hepatocellular carcinoma cells were either cultured in an interwoven hollow fiber bioreactor or on a conventional two-dimensional (2D) surface. The cells were harvested and RNA sequencing was performed to identify the differentially expressed genes between the two culture conditions. Ingenuity Pathway Analysis (IPA) software was utilized to algorithmically generate gene networks and canonical pathways that were either activated or repressed as a result of the experimental conditions. Results: RNA sequencing analysis identified 5,711 genes that were differentially expressed (FDR < 0.05). Of these, 1,248 genes were biologically upregulated (FC > 2) and 629 genes were biologically downregulated (FC < -2). Several of these upregulated genes have known associations with metabolic activity, cellular proliferation, and apico-basal cell organization. IPA analysis confirmed the likely activation of metabolic pathways in 3D culture and suggest a strong relationship between hepatic metabolism, cell-cell interactions, and polarity markers Conclusions: Our results demonstrate the likely activation of metabolic pathways in 3D culture and suggest a strong relationship between hepatic metabolism, cell-cell interactions, and polarity markers. Further experimentation must be performed to further elucidate the connections between these pathways.

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Project description:Liver-on-a-chip models predictive for both metabolism and transport of drug candidates in humans are lacking. Here, we have established an advanced, bioengineered and animal-free hepatocyte-like millifluidic system based on 3D hollow fiber membranes (HFMs), recombinant human laminin 332 and adult human stem cell-derived organoids. Organoid cells formed polarized and tight monolayers on HFMs, which displayed improved hepatocyte-like maturation over standard organoid cultures in Matrigel from matched donors. mRNA sequencing and immunofluorescence revealed that HFM cultures exhibited the expression of broad panel of phase I and II drug metabolizing enzymes, and of drug transporters. Functionally, in both static conditions and under flow circulation, HFM monolayers metabolized a cocktail of drugs that are targets for most important phase I and II enzymes. In addition, we were able to study disposition of those parental compounds and their metabolites in the basal circulation and apical compartment of the chip. Moreover, we demonstrated that our system can be used to study drug disposition in a modular setting with other PK/ADME-relevant organ systems. In conclusion, we have generated a proof-of-concept liver organoid-on-a-chip model for examining both metabolism and transport of drugs, which can be further developed towards prediction of PK/ADME profiles in humans.

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