Global gene expression changes including drug metabolism and disposition induced by three-dimensional culture of HepG2 cells-Involvement of microtubules

Horiuchi, Shigetomo; Ishida, Susumu; Hongo, Tomokatsu; Ishikawa, Y.; Miyajima, Atsuko; Sawada, Jun‐ichi; Ohno, Yasuo; Nakazawa, Ken; Ozawa, Shogo

doi:10.1016/j.bbrc.2008.11.088

Cited by 21 publications

(11 citation statements)

References 20 publications

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“…This response, as well as the organization of the cytoskeleton in the first few hours via integrin signaling, could lead to enhancement of the relevant renal functions. Several groups have already reported that microtubule stabilization modulates the expression of a number of genes, including drug metabolism‐related genes 34, 35. Finally, the MDCK were submitted to continuous flow for 72 h in the biochip cultures.…”

Section: Discussionmentioning

confidence: 99%

Analysis of transcriptomic and proteomic profiles demonstrates improved Madin–Darby canine kidney cell function in a renal microfluidic biochip

Snouber

Letourneur

Chafey

et al. 2011

Biotechnology Progress

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We have evaluated the influence of the microfluidic environment on renal cell functionality. For that purpose, we performed a time lapse transcriptomic and proteomic analysis in which we compared gene and protein expressions of Madin-Darby canine kidney cells after 24 h and 96 h of culture in both microfluidic biochips and plates. The transcriptomic and proteomic integration revealed that the ion transporters involved in calcium, phosphate, and sodium homoeostasis and several genes involved in H(+) transporters and pH regulation were up-regulated in microfluidic biochips. Concerning drug metabolism, we found Phase I (CYP P450), Phase II enzymes (GST), various multidrug resistance genes (MRP), and Phase III transporters (SLC) were also up-regulated in the biochips. Furthermore, the study shows that those inductions were correlated with the induction of the Ahr and Nrf-2 dependent pathways, which results in a global cytoprotective response induced by the microenvironment. However, there was no apoptosis situation or cell death in the biochips. Microfluidic biochips may thus provide an important insight into exploring xenobiotic injury and transport modifications in this type of bioartificial microfluidic kidney. Finally, the investigation demonstrated that combining the transcriptomic and proteomic analyses obtained from a cell "on chip" culture would provide a pertinent new tool in the mechanistic interpretation of cellular mechanisms for predicting kidney cell toxicity and renal clearance in vitro.

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Section: Discussionmentioning

confidence: 99%

Analysis of transcriptomic and proteomic profiles demonstrates improved Madin–Darby canine kidney cell function in a renal microfluidic biochip

Snouber

Letourneur

Chafey

et al. 2011

Biotechnology Progress

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“…As former work with the human liver-derived cell line HepG2 in 3D cell culture configuration revealed an increased expression of liverspecific genes compared to monolayer culture (Altmann et al 2008;Horiuchi et al 2009), we used HepG2 cells as in vitro model to study the hepatocellular metabolic functions in our microstructured 3D substrates and to examine the suitability of the culture system for screening purposes. In addition, we compared our system with a commercially available 3D culture system based on porous alginate sponges in 96-well plates.…”

Section: Discussionmentioning

confidence: 99%

Microstructuring of multiwell plates for three-dimensional cell culture applications by ultrasonic embossing

Altmann

Ahrens

Welle

et al. 2011

Biomed Microdevices

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Since three-dimensional (3D) cell culture models better reflect tissues in vivo in terms of cell shape and microenvironment compared to conventional monolayer cultures, 3D tissue culture substrates gain more importance for a wide range of biological applications like drug discovery, toxicological studies, cancer and stem cell research. In this study we developed a method for the fabrication of 3D cell culture substrates in a multiwell plate format by microstructuring the bottom of 96-well cell culture plates using an ultrasonic embossing process. The resulting microstructured area consists of cubic microcavities in which adherent multicellular aggregates can be formed. We performed the biological evaluation of the system with the liver-derived human cell-line HepG2 and compared the novel substrate with a commercially available 3D culture system comprising porous alginate sponges. Metabolic activity (alamarBlue® reduction) and induction of four biotransformation enzymes (EROD, ECOD, UGT, SULT) were determined by fluorimetry or HPLC. Our results revealed that HepG2 cells in microstructured plates showed a higher mitochondrial activity, as well as enzyme activity of ECOD and UGT after treatment with an inducer when compared to cells cultured in alginate sponges at otherwise comparable conditions. Since we have modified standard cell culture plates, the obtained system is adaptable to automated screening and might be useful for all kinds of cultures including adult, progenitor and stem cells which need a 3D culture configuration to restore or maintain the differentiated status.

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“…These cell-based assays are designed to approximate the drug metabolism that takes place in the liver, and allow for the identification of direct toxicity and the toxicity of drug metabolites. The most widely used in vitro toxicity screening systems are HepG2 liver carcinoma cells, microsomes from cadaveric liver, and primary hepatocytes from surgeries or cadavers [20][21][22][23][24][25][26][27][28][29][30].…”

Section: Reviewmentioning

confidence: 99%

Ethnically diverse pluripotent stem cells for drug development

Fakunle

Loring

2012

Trends in Molecular Medicine

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Global gene expression changes including drug metabolism and disposition induced by three-dimensional culture of HepG2 cells-Involvement of microtubules

Cited by 21 publications

References 20 publications

Analysis of transcriptomic and proteomic profiles demonstrates improved Madin–Darby canine kidney cell function in a renal microfluidic biochip

Analysis of transcriptomic and proteomic profiles demonstrates improved Madin–Darby canine kidney cell function in a renal microfluidic biochip

Microstructuring of multiwell plates for three-dimensional cell culture applications by ultrasonic embossing

Ethnically diverse pluripotent stem cells for drug development

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