Modulation of cytochrome P450 gene expression in primary hepatocytes on various artificial extracellular matrices

Adachi, Tatsuhiko; Goto, Mitsuaki; Cho, Chong‐Su; Akaike, Toshihiro

doi:10.1016/j.bbrc.2011.09.004

Cited by 4 publications

(5 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Many of the studies have used primary cells and these have shown that 3D culture prolongs crucial liver-like functionality in primary rat (Brophy et al , 2009; Schutte et al , 2011), porcine (Nelson et al , 2010), and human hepatocytes (Tostoes et al , 2012). Spheroid formation maintains cytochrome P450 expression (Adachi et al , 2011) enhances drug efflux activities (Oshikata et al , 2011) and preserves (compared with adult liver) gene expression of several key hepatic markers (including adhesion molecules, transcription factors, metabolic enzymes including cytochrome P450, and transporters) (Sakai et al , 2010). Although most studies have used primary hepatocytes, there are several reports in the literature that illustrate that immortal hepatocytes also can recover crucial liver-like functionality and that this might increase their value for toxicological studies (Elkayam et al , 2006; Selden et al , 2000; Xu et al , 2003a,b).…”

mentioning

confidence: 99%

Determination of Drug Toxicity Using 3D Spheroids Constructed From an Immortal Human Hepatocyte Cell Line

Fey

Wrzesinski

2012

Toxicological Sciences

166

157

View full text Add to dashboard Cite

Numerous publications have documented that the immortal cells grown in three-dimensional (3D) cultures possess physiological behavior, which is more reminiscent of their parental organ than when the same cells are cultivated using classical two-dimensional (2D) culture techniques. The goal of this study was to investigate whether this observation could be extended to the determination of LD50 values and whether 3D data could be correlated to in vivo observations. We developed a noninvasive means to estimate the amount of protein present in a 3D spheroid from it is planar area (± 21%) so that a precise dose can be provided in a manner similar to in vivo studies. This avoided correction of the actual dose given based on a protein determination after treatment (when some cells may have lysed). Conversion of published in vitro LC50 data (mM) for six common drugs (acetaminophen, amiodarone, diclofenac, metformin, phenformin, and valproic acid) to LD50 data (mg compound/mg cellular protein) showed that the variation in LD50 values was generally less than that suggested by the original LC50 data. Toxicological analysis of these six compounds in 3D spheroid culture (either published or presented here) demonstrated similar LD50 values. Although in vitro 2D HepG2 data showed a poor correlation, the primary hepatocyte and 3D spheroid data resulted in a much higher degree of correlation with in vivo lethal blood plasma levels. These results corroborate that 3D hepatocyte cultures are significantly different from 2D cultures and are more representative of the liver in vivo.

show abstract

mentioning

confidence: 99%

Determination of Drug Toxicity Using 3D Spheroids Constructed From an Immortal Human Hepatocyte Cell Line

Fey

Wrzesinski

2012

Toxicological Sciences

166

157

View full text Add to dashboard Cite

show abstract

“…The ECM of liver tissue is not only composed of collagen, but also many other kinds of proteins as typified by laminin, fibronectin, proteoglycan, and so on. Similarly, Adachi et al reported that the mRNA expression of the CYP gene in hepatocytes cultured on lactose‐carrying styrene polymer‐coated dishes was significantly higher compared to collagen‐coated dishes . In addition, Hoshiba et al also reported that the hepatocytes grown on PVLA‐coated dish were capable to maintain several liver‐specific functions …”

Section: Discussionmentioning

confidence: 99%

“…Similarly, Adachi et al reported that the mRNA expression of the CYP gene in hepatocytes cultured on lactose-carrying styrene polymer-coated dishes was significantly higher compared to collagen-coated dishes. 30 In addition, Hoshiba et al also reported that the hepatocytes grown on PVLA-coated dish were capable to maintain several liver-specific functions. 31 PVLA is one of the specific polymers that adheres hepatocytes through the interaction between the asialoglycoprotein receptor (ASGPR) of hepatocytes and the galactose moieties of PVLA polymer.…”

Section: Discussionmentioning

confidence: 99%

“…Matrix coating was carried out as previously described . Briefly, 3 mg/mL collagen, 100 μg/mL PVLA, and 0.5 mg/mL or 5 mg/mL GA solution were added to a 24‐well plate at 37°C and incubated overnight.…”

Section: Methodsmentioning

confidence: 99%

“…Matrix coating was carried out as previously described. [28][29][30][31][32] Briefly, 3 mg/mL collagen, 100 lg/mL PVLA, and 0.5 mg/mL or 5 mg/mL GA solution were added to a 24-well plate at 378C and incubated overnight. After incubation, the nonabsorbed solution was removed and washed with PBS three times.…”

Section: Preparation Of the Extracellular Matrix-coated Dishmentioning

confidence: 99%

See 2 more Smart Citations

Fabrication of 3D‐culture platform with sandwich architecture for preserving liver‐specific functions of hepatocytes using 3D bioprinter

Arai

Yoshida

Okabe

et al. 2017

J Biomedical Materials Res

Self Cite

View full text Add to dashboard Cite

The development of new three-dimensional (3D) cell culture system that maintains the physiologically relevant signals of hepatocytes is essential in drug discovery and tissue engineering research. Conventional two-dimensional (2D) culture yields cell growth, proliferation, and differentiation. However, gene expression and signaling profiles can be different from in vivo environment. Here, we report the fabrication of a 3D culture system using an artificial scaffold and our custom-made inkjet 3D bioprinter as a new strategy for studying liver-specific functions of hepatocytes. We built a 3D culture platform for hepatocytes-attachment and formation of cell monolayer by interacting the galactose chain of galactosylated alginate gel (GA-gel) with asialoglycoprotein receptor (ASGPR) of hepatocytes. The 3D geometrical arrangement of cells was controlled by using 3D bioprinter, and cell polarity was controlled with the galactosylated hydrogels. The fabricated GA-gel was able to successfully promote adhesion of hepatocytes. To observe liver-specific functions and to mimic hepatic cord, an additional parallel layer of hepatocytes was generated using two gel sheets. These results indicated that GA-gel biomimetic matrices can be used as a 3D culture system that could be effective for the engineering of liver tissues. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1583-1592, 2017.

show abstract

Three‐Dimensional Culture Systems and Humanized Liver Models Using Hepatic Stem Cells for Enhanced Toxicity Assessment

Zhang

Zheng

Taniguchi

2016

Stem Cells in Toxicology and Medicine

View full text Add to dashboard Cite

Modulation of cytochrome P450 gene expression in primary hepatocytes on various artificial extracellular matrices

Cited by 4 publications

References 21 publications

Determination of Drug Toxicity Using 3D Spheroids Constructed From an Immortal Human Hepatocyte Cell Line

Determination of Drug Toxicity Using 3D Spheroids Constructed From an Immortal Human Hepatocyte Cell Line

Fabrication of 3D‐culture platform with sandwich architecture for preserving liver‐specific functions of hepatocytes using 3D bioprinter

Three‐Dimensional Culture Systems and Humanized Liver Models Using Hepatic Stem Cells for Enhanced Toxicity Assessment

Contact Info

Product

Resources

About