Isolation of Mouse Hepatocytes

Edwards, Meredith; Houseman, Lyndsey; Phillips, Ian; Shephard, Elizabeth A.

doi:10.1007/978-1-62703-321-3_24

Cited by 22 publications

(14 citation statements)

References 6 publications

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“…Importantly, ATPIF1 -/- hepatocytes had increased cell viability relative to WT hepatocytes following treatment with antimycin (Figure 4D), which demonstrates that the beneficial effects of ATPIF1 loss under severe ETC dysfunction are not limited to rapidly proliferating cancer cell lines, but can also occur in post-mitotic, differentiated cells that better recapitulate the metabolism of tissues affected in severe mitochondrial respiratory chain disorders (Vander Heiden et al, 2009). The smaller effects of ATPIF1 loss on hepatocyte viability during antimycin treatment, as compared to KBM7 cells, are partially due to the frailty of primary mouse hepatocytes when cultured ex vivo (Edwards et al, 2013, Klaunig et al, 1981). Taken together, these data demonstrate that ATPIF1 loss in primary hepatocytes can ameliorate the effects of complex III blockade.…”

Section: Resultsmentioning

confidence: 99%

Inhibition of ATPIF1 Ameliorates Severe Mitochondrial Respiratory Chain Dysfunction in Mammalian Cells

et al. 2014

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SUMMARY Mitochondrial respiratory chain disorders are characterized by loss of electron transport chain (ETC) activity. Although the causes for many such diseases are known, there is a lack of effective therapies. To identify genes that confer resistance to severe ETC dysfunction when inactivated, we performed a genome-wide genetic screen in haploid human cells with the mitochondrial complex III inhibitor, antimycin. This screen revealed that loss of ATPIF1 strongly protects against antimycin-induced ETC dysfunction and cell death by allowing for maintenance of mitochondrial membrane potential. ATPIF1 loss protects against other forms of ETC dysfunction and is even essential for the viability of human ρ0 cells lacking mitochondrial DNA, a common system for studying ETC dysfunction. Importantly, inhibition of ATPIF1 ameliorates complex III blockade in primary hepatocytes, a cell type afflicted in severe mitochondrial disease. Taken together, these results suggest that inhibition of ATPIF1 can ameliorate severe ETC dysfunction in mitochondrial pathology.

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Section: Resultsmentioning

confidence: 99%

Inhibition of ATPIF1 Ameliorates Severe Mitochondrial Respiratory Chain Dysfunction in Mammalian Cells

et al. 2014

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“…Mouse primary hepatocytes were isolated as described previously [28] , [29] . In brief, mice were anaesthetized by intraperitoneal injection of 50 mg/kg pentobarbital.…”

Section: Methodsmentioning

confidence: 99%

Hepatic Carboxylesterase 1 Is Induced by Glucose and Regulates Postprandial Glucose Levels

Yin

et al. 2014

PLoS ONE

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Metabolic syndrome, characterized by obesity, hyperglycemia, dyslipidemia and hypertension, increases the risks for cardiovascular disease, diabetes and stroke. Carboxylesterase 1 (CES1) is an enzyme that hydrolyzes triglycerides and cholesterol esters, and is important for lipid metabolism. Our previous data show that over-expression of mouse hepatic CES1 lowers plasma glucose levels and improves insulin sensitivity in diabetic ob/ob mice. In the present study, we determined the physiological role of hepatic CES1 in glucose homeostasis. Hepatic CES1 expression was reduced by fasting but increased in diabetic mice. Treatment of mice with glucose induced hepatic CES1 expression. Consistent with the in vivo study, glucose stimulated CES1 promoter activity and increased acetylation of histone 3 and histone 4 in the CES1 chromatin. Knockdown of ATP-citrate lyase (ACL), an enzyme that regulates histone acetylation, abolished glucose-mediated histone acetylation in the CES1 chromatin and glucose-induced hepatic CES1 expression. Finally, knockdown of hepatic CES1 significantly increased postprandial blood glucose levels. In conclusion, the present study uncovers a novel glucose-CES1-glucose pathway which may play an important role in regulating postprandial blood glucose levels.

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“…Hepatocytes were isolated according to the method described by (Edwards et al 2013), in which mice anesthetized by inhalation of isoflurane were subjected to liver perfusion technique in situ (de Morais et al 2012) with collagenase. After perfusion, the liver was filtered and the hepatocytes obtained were counted by TRIPAN Blue exclusion method.…”

Section: Isolation and Culture Of Hepatocytesmentioning

confidence: 99%

Short-term treatment with metformin reduces hepatic lipid accumulation but induces liver inflammation in obese mice

et al. 2018

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The study aimed to evaluate the metabolic and inflammatory effects of short-term treatments (10 days) with metformin (MET) on the NAFLD caused by a high-fat diet (HFD) in C57BL/6 mice. After the treatment, histological liver slices were obtained, hepatocytes and macrophages were extracted and cultured with phosphate buffered saline, LPS (2.5 µg/mL) and MET (1 µM) for 24 h. Cytokine levels were determined by ELISA. NAFLD caused by the HFD was partially reduced by MET. The lipid accumulation induced by the HFD was not associated with liver inflammation; however, MET seemed to promote pro-inflammatory effects in liver, since it increased hepatic concentration of IL-1β, TNF-α, IL-6, MCP-1 and IFN-γ. Similarly, MET increased the concentration of IL-1β, IL-6 in hepatocyte cultures. However, in macrophages culture, MET lowered levels of IL-1β, IL-6 and TNF-α stimulated by LPS. Overall, MET reduced liver NAFLD but promoted hepatocyte increase in pro-inflammatory cytokines, thus, leading to liver inflammation.

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Isolation of Mouse Hepatocytes

Cited by 22 publications

References 6 publications

Inhibition of ATPIF1 Ameliorates Severe Mitochondrial Respiratory Chain Dysfunction in Mammalian Cells

Inhibition of ATPIF1 Ameliorates Severe Mitochondrial Respiratory Chain Dysfunction in Mammalian Cells

Hepatic Carboxylesterase 1 Is Induced by Glucose and Regulates Postprandial Glucose Levels

Short-term treatment with metformin reduces hepatic lipid accumulation but induces liver inflammation in obese mice

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