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Novel indirect co-culture of immortalized hepatocytes with monocyte derived macrophages is supported by pro-inflammatory cytokine networks

ABSTRACT: Novel indirect co-culture of immortalized hepatocytes with monocyte derived macrophages is supported by pro-inflammatory cytokine networks

PROVIDER: PRJNA658539 | ENA |

REPOSITORIES: ENA

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Novel indirect co-culture of immortalized hepatocytes with monocyte derived macrophages is supported by pro-inflammatory cytokine networks

Project description:The liver is composed of different cell populations. Interactions of different cell populations can be investigated bya newly established indirect co-culture system consisting of immortalised primary human hepatocytes and human monocyte derived macrophages(MDMs). Using the time-dependent cytokine secretion of the co-cultures and singlecultures, correlation networks(includingthe cytokinesG-CSF, CCL3, MCP-1, CCL20, FGF, TGF-β1, GM-CSF, IL-8 IL-6, IL-1β, and IL-18)were generated and the correlationswere validatedbyapplication of IL-8and TNF-α-neutralising antibodies. Thedata reveal that IL-8 is crucial for the interaction between hepatocytes and macrophages in vitro.In addition, transcriptome analyses showedthat a change in the ratio between macrophages and hepatocytes may triggerpro-inflammatory signalling pathways of the acute phase responseand the complement system(release of, e.g., certaincyto-and chemokines).Using diclofenac and LPS showed that the release of cytokines is increasingwith higher ratios of MDMs. Altogether we coulddemonstratethat the current co-culture system is better suited to mirror thein vivosituation when compared to previously established co-culturesystems composed of HepG2and differentiated THP-1cells.Further, our data reveal that the cytokine IL-8 is crucial for the interaction between hepatocytes and macrophages in vitro.

2021-03-19 | GSE156627 | GEO

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Project description:To investigate the effect of hyperactivation of Notch signaling in hepatocytes, an immortalized human hepatocyte cell line THLE-2 was stably transduced with control vector or plasmid harborning intracellular domain of NOTCH1 (NICD1) and the transcriptomic profiles were analyzed by RNA-seq.

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Project description:Fundamental research and drug development for personalized medicine necessitates cell cultures from defined genetic backgrounds. However, providing sufficient numbers of authentic cells from individuals poses a challenge. Here, we present a new strategy for rapid cell expansion that overcomes current limitations. Using a small gene library, we expanded primary cells from different tissues, donors and species. Cell type specific regimens that allow the reproducible creation of cell lines were identified. In depth characterization of a series of endothelial and hepatocytic cell lines confirmed phenotypic stability and functionality. Applying this technology enables rapid, efficient and reliable production of unlimited numbers of personalized cells. As such, these cell systems support mechanistic studies, epidemiological research and tailored drug development. In these experiments primary murine hepatocytes were compared to immortalized murine hepatocytes with respect to their global gene expression pattern.

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