Determination and Characterization of Tetraspanin-Associated Phosphoinositide-4 Kinases in Primary and Neoplastic Liver Cells

Abstract

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The adenosine monophosphate–activated protein kinase—vacuolar adenosine triphosphatase–pH axis: A key regulator of the profibrogenic phenotype of human hepatic stellate cells

“…Cells were isolated from wedge sections of human liver obtained from patients undergoing liver resections at the Royal Free Hospital after giving informed consent (EC01.14‐RF). Cell isolation was performed according to a published protocol, with modifications for human liver …”

Ammonia produces pathological changes in human hepatic stellate cells and is a target for therapy of portal hypertension

“…Cells were isolated according to Mederacke et al, [30] with the relative modifications for human liver. [31] Briefly, 10 g of total human liver tissue was digested with 0.01% Collagenase, 0.05% Pronase and 0.001% DNase I without performing perfusion. The homogenate was filtered through a 100µm cell strainer and the flow-through was centrifuged at 50xg…”

Reactive gamma-ketoaldehydes as novel activators of hepatic stellate cells in vitro

“…Cells were isolated from resected livers wedges, obtained from patients undergoing surgery at the Royal Free Hospital after giving informed consent (EC01.14-RF). Cells were isolated according to a published protocol [32], with modifications for human liver [2]. In brief, 10 grams of total human liver tissue were digested with 0.01% Collagenase, 0.05% Pronase, and 0.001% DNase I without perfusion.…”

“…However, the chronic activation of this process through persistence of parenchymal injury, causes the progressive accumulation of fibrillar ECM leading to liver cirrhosis [1][2][3]. An established trigger for HSC activation is oxidative stress, and, in particular, the products of lipid oxidation or oxidative modification of membrane polyunsaturated fatty acids.…”

“…Non-enzymatic rearrangement of the bicycloendoperoxide intermediates of these two pathways (PGH 2 or its regioisomers, respectively) yield acyclic -ketoaldehydes. Whilst the free-radical mediated pathway generates a mixture of regio-and stereoisomers, the rearrangement of PGH 2 yields a single stereoisomer of 15-E 2 -IsoLG and 15-D 2 -IsoLG, designated as LGE 2 and LGD 2 respectively [12,13].…”