Ascorbic acid supplementation to primary culture of chicken hepatocytes with non-serum medium

Sasaki, Keisuke; Kitaguchi, Yasunari; Fukuda, Tatsuki; Aoyagi, Yosuke

doi:10.1016/s1357-2725(00)00043-1

Cited by 17 publications

(17 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We chose cultured primary chicken hepatocytes as our experimental model system, because in culture they retained many liver-specific functions and hormone responses in previous biophysiological liver function studies. The primary chicken hepatocyte culture system utilized herein also retains many typical differentiated liver phenotypes, including albumin expression [Hou et al, 2001], p4501A induction [Hou et al, 2001], tyrosine aminotransferase expression [Sasaki et al, 2000], and ascorbate recycling [Sasaki et al, 2001], in an in vitro tissue culture setting. These properties make this system ideal for elucidating the cellular components of L-leucine-mediated hepatocyte proliferation.…”

mentioning

confidence: 99%

L‐leucine increases [³H]‐thymidine incorporation in chicken hepatocytes: Involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways

Lee

et al. 2008

J of Cellular Biochemistry

View full text Add to dashboard Cite

This study examined how L-leucine affected DNA synthesis and cell cycle regulatory protein expression in cultured primary chicken hepatocytes. L-Leucine promoted DNA synthesis in a dose- and time-dependent manner, with concomitant increases in cyclin D1 and cyclin E expression. Phospholipase C (PLC) and protein kinase C (PKC) mediated the L-leucine-induced increases in [3H]-thymidine incorporation and cyclin D1/CDK4 and cyclin E/CDK2 expression, as U73122 (a PLC inhibitor) or bisindolylmaleimide I (a PKC blocker) inhibited these effects. L-Leucine also increased PKC phosphorylation and intracellular Ca2+ levels. L-Leucine-mediated increases in [3H]-thymidine incorporation and cyclin/CDK expression were sensitive to LY 294002 (PI3K inhibitor), Akt inhibitor, PD 98059 (MEK inhibitor). It was also observed that L-leucine-induced increases of cyclin/CDK expression were inhibited by PI3K siRNA and ERK siRNA; L-leucine increased extracellular signal-regulated kinases 1/2 (ERK1/2) and Akt phosphorylation levels. Bisindolylmaleimide I attenuated L-leucine-induced phosphorylation of ERK1/2 but did not influence Akt phosphorylation, and PI3K siRNA and LY 294002 inhibited L-leucine-induced ERK1/2 phosphorylation, suggesting some cross-talk between the PKC and ERK1/2 or PI3K/Akt and ERK1/2 pathways. L-Leucine also increased the levels of phosphorylated molecular target of rapamycin (mTOR) and two of its targets, ribosomal protein S6 kinase (p70S6K), and 4E binding protein 1 (4E-BP1); furthermore, rapamycin (an mTOR inhibitor) blocked all of the mitogenic effects of L-leucine. In addition, Akt inhibitor blocked L-leucine-induced mTOR phosphorylation. In conclusion, L-leucine stimulated DNA synthesis and promoted cell cycle progression in primary cultured chicken hepatocytes through PKC, ERK1/2, PI3K/Akt, and mTOR.

show abstract

mentioning

confidence: 99%

L‐leucine increases [³H]‐thymidine incorporation in chicken hepatocytes: Involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways

Lee

et al. 2008

J of Cellular Biochemistry

View full text Add to dashboard Cite

show abstract

“…Primary hepatocyte cultures have been used in many biophysiological studies of liver function because they retain many liver-specific functions and respond to various hormones through the expression of liver-specific functions (Lee et al, 2006;Suh et al, 2008). The primary chicken hepatocytes culture system used in this study also retains the in vitro differentiated phenotype that is typical of the liver, including albumin expression (Hou et al, 2001), P450 1A induction (Hou et al, 2001), tyrosine aminotransferase expression (Sasaki et al, 2000), and ascorbate recycling (Sasaki et al, 2001). Therefore, in the present study, we investigated the effect of IL-6 on 2-DG uptake and related signaling pathways in primary cultured hepatocytes.…”

mentioning

confidence: 99%

Interleukin‐6 promotes 2‐deoxyglucose uptake through p44/42 MAPKs activation via Ca²⁺/PKC and EGF receptor in primary cultured chicken hepatocytes

Lee

Han

2008

Journal Cellular Physiology

View full text Add to dashboard Cite

Interleukin-6 (IL-6) is involved in a variety of biological responses, including the glucose metabolism and cell growth, which is a critical physiological function requiring multiple metabolic pathways. Therefore, in the present study, we examined the effect of IL-6 on 2-deoxyglucose (2-DG) uptake and the related signaling pathways in primary cultured chicken hepatocytes. IL-6 increased 2-DG uptake in a time- (> or =4 h) and a dose -(> or =5 ng/ml) dependent manner. Indeed, IL-6 increased GLUT-2 mRNA and protein expression as well as 2-DG uptake, which were blocked by actinomycin D (AD, transcription inhibitor) and cycloheximide (CHX, translation inhibitor). IL-6 (10 ng/ml) increased the level of IL-6Ralpha and glycoprotein (gp) 130 (IL-6Rbeta) protein expressions. IL-6 increased Janus Kinase (JAK)-2, signal transducer and activator of transcription (STAT)-3 phosphorylation, intracellular Ca(2+) concentration, and PKC phosphorylation. IL-6-induced increase of 2-DG uptake and GLUT-2 protein expression were blocked by JAK2-specific siRNA, a STAT3 inhibitor, staurosporine, and bisindolylmaleimide I (PKC inhibitors). In addition, IL-6 increased EGFR/src/FAK, PI3K/Akt phosphorylation and 2-DG uptake as well as GLUT-2 protein expression, which were blocked by AG 1478 (EGF receptor inhibitor), PP2 (src family of tyrosine kinase inhibitor), PI3K-specific siRNA, and a Akt inhibitor. Furthermore, IL-6 increased p44/42 MAPKs phosphorylation and p44 and p42 MAPK-specific siRNA mixture blocked IL-6-induced increase of 2-DG uptake and GLUT-2 protein expression. In conclusion, IL-6 stimulates the 2-DG uptake through p44/42 MAPKs activation via Ca(2+)/PKC and EGF receptor in primary cultured chicken hepatocytes.

show abstract

“…Because of their easy access, the cells are widely utilized in clinical trials to investigate hepatic function, estimate xenobiotic metabolism, establish artificial organs in vitro, and further regenerate tissue in vivo. 2,3 However, hepatocytes tend to lose their reproductive activity and specific liver function steadily after isolation. Meanwhile, attempts to replace primary cells by using permanent cells or even cancer cell lines have also encountered drawbacks because the substitute cells are not fully consistent with primary cells with regard to metabolism and functionality.…”

Section: Introductionmentioning

confidence: 99%

A hybrid substratum for primary hepatocyte culture that enhances hepatic functionality with low serum dependency

Meng¹,

Tao²,

Qiu³

et al. 2015

IJN

View full text Add to dashboard Cite

Cell culture systems have proven to be crucial for the in vitro maintenance of primary hepatocytes and the preservation of hepatic functional expression at a high level. A poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide) matrix can recognize cells and promote liver function in a spheroid structure because of a specific galactose–asialoglycoprotein receptor interaction. Meanwhile, a fusion protein, E-cadherin-Fc, when incubated with various cells, has shown an enhancing effect on cellular viability and metabolism. Therefore, a hybrid substratum was developed for biomedical applications by using both of these materials to combine their advantages for primary hepatocyte cultures. The isolated cells showed a monolayer aggregate morphology on the coimmobilized surface and displayed higher functional expression than cells on traditional matrices. Furthermore, the hybrid system, in which the highest levels of cell adhesion and hepatocellular metabolism were achieved with the addition of 1% fetal bovine serum, showed a lower serum dependency than the collagen/gelatin-coated surface. Accordingly, this substrate may attenuate the negative effects of serum and further contribute to establishing a defined culture system for primary hepatocytes.

show abstract

Ascorbic acid supplementation to primary culture of chicken hepatocytes with non-serum medium

Cited by 17 publications

References 17 publications

L‐leucine increases [³H]‐thymidine incorporation in chicken hepatocytes: Involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways

L‐leucine increases [³H]‐thymidine incorporation in chicken hepatocytes: Involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways

Interleukin‐6 promotes 2‐deoxyglucose uptake through p44/42 MAPKs activation via Ca²⁺/PKC and EGF receptor in primary cultured chicken hepatocytes

A hybrid substratum for primary hepatocyte culture that enhances hepatic functionality with low serum dependency

Contact Info

Product

Resources

About

Ascorbic acid supplementation to primary culture of chicken hepatocytes with non-serum medium

Cited by 17 publications

References 17 publications

L‐leucine increases [3H]‐thymidine incorporation in chicken hepatocytes: Involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways

L‐leucine increases [3H]‐thymidine incorporation in chicken hepatocytes: Involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways

Interleukin‐6 promotes 2‐deoxyglucose uptake through p44/42 MAPKs activation via Ca2+/PKC and EGF receptor in primary cultured chicken hepatocytes

A hybrid substratum for primary hepatocyte culture that enhances hepatic functionality with&nbsp;low serum dependency

Contact Info

Product

Resources

About

L‐leucine increases [³H]‐thymidine incorporation in chicken hepatocytes: Involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways

L‐leucine increases [³H]‐thymidine incorporation in chicken hepatocytes: Involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways

Interleukin‐6 promotes 2‐deoxyglucose uptake through p44/42 MAPKs activation via Ca²⁺/PKC and EGF receptor in primary cultured chicken hepatocytes

A hybrid substratum for primary hepatocyte culture that enhances hepatic functionality with low serum dependency