A new culture system activating hepatic function of HepG2 cells utilizing a collagen vitrigel membrane chamber and its application to liver metabolism and toxicity assays

AbstractWound healing is regulated by complex interactions between the keratinocytes and other cell types including fibroblasts. Recently, adipose-derived mesenchymal stromal/stem cells (ASCs) have been reported to influence wound healing positively via paracrine involvement. However, their roles in keratinocytes are still obscure. Therefore, investigation of the precise effects of ASCs on keratinocytes in an in vitro culture system is required. Our recent data indicate that the epidermal equivalents became thicker on a collagen vitrigel membrane co-cultured with human ASCs (hASCs). Co-culturing the human primary epidermal keratinocytes (HPEK) with hASCs on a collagen vitrigel membrane enhanced their abilities for cell proliferation and adhesion to the membrane but suppressed their differentiation suggesting that hASCs could maintain the undifferentiated status of HPEK. Contrarily, the effects of co-culture using polyethylene terephthalate or polycarbonate membranes for HPEK were completely opposite. These differences may depend on the protein permeability and/or structure of the membrane. Taken together, our data demonstrate that hASCs could be used as a substitute for fibroblasts in skin wound repair, aesthetic medicine, or tissue engineering. It is also important to note that a co-culture system using the collagen vitrigel membrane allows better understanding of the interactions between the keratinocytes and ASCs.

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UPLC-MS/MS analysis of ochratoxin A metabolites produced by Caco-2 and HepG2 cells in a co-culture system

Food and Chemical Toxicology ◽

10.1016/j.fct.2017.09.011 ◽

2017 ◽

Vol 109 ◽

pp. 333-340 ◽

Cited By ~ 7

Author(s):

Cyndia A. González-Arias ◽

Sonia Marín ◽

Aurora E. Rojas-García ◽

Vicente Sanchis ◽

Antonio J. Ramos

Keyword(s):

Ochratoxin A ◽

Hepg2 Cells ◽

Culture System ◽

Ms Analysis

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P120 - In-vitro evaluation of primary animal hepatic function using a 3D-spheroid culture system

Drug Metabolism and Pharmacokinetics ◽

10.1016/j.dmpk.2020.04.121 ◽

2020 ◽

Vol 35 (1) ◽

pp. S59

Author(s):

Julia Tritapoe ◽

Sujoy Lahiri ◽

Michael Millett ◽

Theresa Nguyen ◽

Mark Powers ◽

...

Keyword(s):

Culture System ◽

Hepatic Function ◽

Spheroid Culture ◽

In Vitro Evaluation

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HepG2-NIAS cells, a new subline of HepG2 cells that can enhance not only CYP3A4 activity but also expression of drug transporters and form bile canaliculus-like networks by the oxygenation culture via a collagen vitrigel membrane

The Journal of Toxicological Sciences ◽

10.2131/jts.47.39 ◽

2022 ◽

Vol 47 (1) ◽

pp. 39-50

Author(s):

Toshiaki Takezawa ◽

Miaki Uzu

Keyword(s):

Hepg2 Cells ◽

Drug Transporters ◽

Bile Canaliculus ◽

Cyp3a4 Activity ◽

Collagen Vitrigel

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A long-term culture system based on a collagen vitrigel membrane chamber that supports liver-specific functions of hepatocytes isolated from mice with humanized livers

The Journal of Toxicological Sciences ◽

10.2131/jts.43.521 ◽

2018 ◽

Vol 43 (8) ◽

pp. 521-529 ◽

Cited By ~ 12

Author(s):

Ryuji Watari ◽

Motoharu Kakiki ◽

Ayumi Oshikata ◽

Toshiaki Takezawa ◽

Chihiro Yamasaki ◽

...

Keyword(s):

Culture System ◽

Long Term Culture ◽

Collagen Vitrigel

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Pro-inflammatory signaling by 24,25-dihydroxyvitamin D3 in HepG2 cells

Journal of Molecular Endocrinology ◽

10.1530/jme-16-0009 ◽

2016 ◽

Vol 57 (2) ◽

pp. 87-96 ◽

Cited By ~ 6

Author(s):

Kent Wehmeier ◽

Luisa M Onstead-Haas ◽

Norman C W Wong ◽

Arshag D Mooradian ◽

Michael J Haas

Keyword(s):

Vitamin D ◽

Signaling Pathways ◽

Hepg2 Cells ◽

Density Lipoprotein ◽

Hepatic Function ◽

Peroxisome Proliferator ◽

Dihydroxyvitamin D3 ◽

Peroxisome Proliferator Activated Receptor ◽

Receptor Alpha ◽

Hydroxyvitamin D

The vitamin D metabolite 24,25-dihydroxyvitamin D3(24, 25[OH]2D3) was shown to induce nongenomic signaling pathways in resting zone chondrocytes and other cells involved in bone remodeling. Recently, our laboratory demonstrated that 24,25-[OH]2D3but not 25-hydroxyvitamin D3, suppresses apolipoprotein A-I (apo A-I) gene expression and high-density lipoprotein (HDL) secretion in hepatocytes. Since 24,25-[OH]2D3has low affinity for the vitamin D receptor (VDR) and little is known with regard to how 24,25-[OH]2D3modulates nongenomic signaling in hepatocytes, we investigated the capacity of 24,25-[OH]2D3to activate various signaling pathways relevant to apo A-I synthesis in HepG2 cells. Treatment with 24,25-[OH]2D3resulted in decreased peroxisome proliferator-activated receptor alpha (PPARα) expression and retinoid-X-receptor alpha (RXRα) expression. Similarly, treatment of hepatocytes with 50 nM 24,25-[OH]2D3for 1–3 h induced PKCα activation as well as c-jun-N-terminal kinase 1 (JNK1) activity and extracellular-regulated kinase 1/2 (ERK1/2) activity. These changes in kinase activity correlated with changes in c-junphosphorylation, an increase in AP-1-dependent transcriptional activity, as well as repression of apo A-I promoter activity. Furthermore, treatment with 24,25-[OH]2D3increased IL-1β, IL-6, and IL-8 expression by HepG2 cells. These observations suggest that 24,25-[OH]2D3elicits several novel rapid nongenomic-mediated pro-inflammatory protein kinases targeting AP1 activity, increasing pro-inflammatory cytokine expression, potentially impacting lipid metabolism and hepatic function.

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Butyric Acid Added Apically to Intestinal Caco-2 Cells Elevates Hepatic ApoA-I Transcription and Rescues Lower ApoA-I Expression in Inflamed HepG2 Cells Co-Cultured in the Basolateral Compartment

Biomolecules ◽

10.3390/biom11010071 ◽

2021 ◽

Vol 11 (1) ◽

pp. 71

Author(s):

Jehad Z. Tayyeb ◽

Herman E. Popeijus ◽

Ronald P. Mensink ◽

Jogchum Plat

Keyword(s):

Butyric Acid ◽

Hepg2 Cells ◽

Culture System ◽

Density Lipoprotein ◽

Short Chain Fatty Acids ◽

Culture Experiment ◽

Apolipoprotein A ◽

Culture Model ◽

Hdl Functionality ◽

Anti Inflammatory

Apolipoprotein A-I (ApoA-I) concentrations are decreased during inflammation, which may reduce high-density lipoprotein (HDL) functionality. Thus, rescuing ApoA-I concentrations during inflammation might help to prevent atherosclerosis. Recent studies have shown that butyric acid (C4) has anti-inflammatory effects and rescues ApoA-I production. However, whether intestinal short chain fatty acids (SCFAs) are able to influence hepatic processes is unknown. Therefore, we investigated C4 anti-inflammatory effects on ApoA-I transcription in the intestine-liver co-culture model. C4 dose-response experiments in the presence or absence of cytokines were performed in a co-culture system including Caco-2 cells, HepG2 cells, or both. Changes in ApoA-I transcription in Caco-2 cells and HepG2 cells were analyzed using qPCR. C4 increased ApoA-I expression in HepG2 cells that cultured alone. When both cells were cultured together, C4 decreased ApoA-I expression in Caco-2 cells and increased ApoA-I expression in HepG2 cells. However, adding C4 to apical Caco-2 cells resulted in a smaller effect in HepG2 cells compared with adding C4 directly to the hepatocytes. Moreover, C4 rescued ApoA-I expression in inflamed HepG2 cells. These findings suggests that intestinal SCFAs can affect hepatic processes. However, the smaller effect in the co-culture experiment indicates cross-talk between intestine and liver.

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