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• W3139174201 abstract "<h3></h3> Liver fibrosis is driven by progressive accumulation of extracellular matrix (ECM), coupled with chronic inflammation. Recent evidence shows that ECM changes in liver disease directly and indirectly modulate the immune system. Since traditional cell culture models often lack immune cell and ECM components, bioengineered models of liver disease using decellularised tissues present an appealing alternative. Here, we describe the generation of a bioengineered model which incorporates the dynamic culture of circulating immune cells with decellularised human liver scaffolds, supported in a custom-made bioreactor to explore interactions between the immune system and liver ECM proteins at different stages of fibrosis. Liver ECM-scaffolds were generated bydecellularisingliver tissue with/without underlying disease from human liver tissue samples from partial hepatectomy, or by perfusion decellularisation of whole rat liver using established protocols. PBMCs isolated from healthy donor whole-blood were first cultured in the bioreactor under semi-continuous perfusion at high (4.22 Pa) or low shear stress (0.34 Pa) and compared to static conditions. Longitudinal profiling of PBMC phenotype was determined byFACs. ECM-scaffold were then perfused with PBMCs and cultured for 5 days. Immune-perfused tissues were stained for PBMC surface markers (T, B, NK cells and macrophages) to identify cell homing. We designed a custom-made bioreactor which can house 6 decellularised human liver scaffolds and which supports dynamic culture of PBMCs. We found that shear stress impacts PBMC viability and that PBMCs are better supported under low shear stress conditions. PBMC phenotype profiling byFACs show no significant changes in proportion of T cells between static and dynamic culture, while an increase in proportion ofTregsand a decrease in proportion of NK cells were detected in dynamic culture. Perfusion of PBMCs in decellularised scaffolds without fibrosis show homing of cells in different areas of the scaffold at all time points, with CD3+ T cells representing 50% of cells and with small clusters of macrophages, monocytes and B cells at 3 days of culture. Dynamic perfusion improved cell density and homing in decellularised liver scaffolds compared to static conditions. Our bioengineered system supports dynamic co-culture of PBMCs and decellularised scaffolds, with varying grades of fibrosis, allowing us to explore the interactions between the ECM and immune cells in liver disease in a dynamic perfusion system. A better understanding of ECM-immune interactions will aid our understanding of the driving factors behind fibrosis and could highlight new disease progression biomarkers and therapeutic targets to treat disease." @default.
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• W3139174201 date "2020-09-01" @default.
• W3139174201 modified "2023-09-26" @default.
• W3139174201 title "P54 Immune-perfusion of liver extracellular matrix-scaffolds in a custom-made bioreactor to explore cell-matrix interaction in liver disease" @default.
• W3139174201 doi "https://doi.org/10.1136/gutjnl-2020-basl.64" @default.
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