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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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Transcription profiling of human HepG2 cells to investigate global gene expression changes including drug metabolism and disposition induced by three-dimensional culture

Project description:We found constitutive upregulation and higher degree induction of drug metabolism and disposition-related genes in a three-dimensional HepG2 culture. The upregulated genes are those believed to be regulated by different regulatory factors. The global gene expression analysis by Affymetrix GeneChip indicated that altered expressions of microtubule-related genes may change expressed levels of drug metabolism and disposition genes. Stabilization of the microtubule molecules with docetaxel, a tubulin stabilizing agent, in the two-dimensional culture showed gene expression patterns similar to those in the three-dimensional culture, indicating that culture environment affects drug metabolism functions in HepG2 cells. Experiment Overall Design: We compared the gene expression data from HepG2 cells cultured in the radial flow bioreactor (RFB) cell culture system for six days to those from HepG2 cells cultured in tissue culture plates for five days. Both cell cultures (RFB culture and plate culture) were performed twice independently. In each culture, three samples were collected from three different portions of the bioreactor or from three different tissue culture plates. Microarray analyses were performed in duplicate for each sample using the Affymetrix human genome U133A GeneChip.

2009-03-08 | E-GEOD-12939 | biostudies-arrayexpress

Global gene expression changes including drug metabolism and disposition induced by three-dimensional culture of HepG2

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Madin-Darby Canine Kidney (MDCK) cells grown in 2-dimensional and 3-dimensional type I collagen culture.

Project description:In order to gain insight into epithelial morphogenesis and the influence of culture geometry on gene expression patterns, Madin Darby Canine Kidney (MDCK) epithelial cells where grown in 2-dimensional (2D) culture or 3-dimensional (3D) culture . MDCK cells cultured in 2D were plated atop a pre-solidified type I collagen gel. Cells cultured under these conditions grew as flat monolayer sheets. In 3D culture, cells are embedded within type I collagen gel. Cells grown under these conditions form large spherical cysts with hollow central lumens. We anticipate, therefore, that these results provide insight into the mechanisms that regulate epithelial cystogenesis.

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Madin-Darby Canine Kidney (MDCK) cells grown in 2-dimensional and 3-dimensional type I collagen culture.

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A hollow fiber membrane-based liver organoid-on-a-chip model for examining drug metabolism and transport

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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Project description:Investigation of whole genome gene expression level changes in hepatocellular carcinoma cell line hepG2 in regular culture, hepG2-slug in regular culture and hepG2-slug on Matrigel. Whole genome gene expression level changes have been compared in hepatocellular carcinoma cell line hepG2 in regular culture, hepG2-slug in regular culture and hepG2-slug on Matrigel. Roche NimbleGen micro-array analysis was employed to assess global genome expression in HepG2 in regular culture, HepG2-slug in regular culture and HepG2-slug on Matrigel. The results demonstrated that the up-regulated genes and the down-regulated genes increased significantly when HepG2-slug cells with VM forming ablity were cultured on Matrigel and formed VM.

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