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Article ; Online: Methods for advanced hepatocyte cell culture in microwells utilizing air bubbles.

Goral, Vasiliy N / Au, Sam H / Faris, Ronald A / Yuen, Po Ki

2015 Volume 15, Issue 4, Page(s) 1032–1037

Abstract: Flat, two-dimensional (2D) cell culture substrates are simple to use but offer little control over cell morphologies and behavior. In this article, we present a number of novel and unique methods for advanced cell culture in microwells utilizing air ... ...

Abstract	Flat, two-dimensional (2D) cell culture substrates are simple to use but offer little control over cell morphologies and behavior. In this article, we present a number of novel and unique methods for advanced cell culture in microwells utilizing air bubbles as a way to seed cells in order to provide substantial control over cellular microenvironments and organization to achieve specific cell-based applications. These cell culture methods enable controlled formation of stable air bubbles in the microwells that spontaneously formed when polar solvents such as cell culture media are loaded. The presence of air bubbles (air bubble masking) enables highly controllable cell patterning and organization of seeded cells as well as cell co-culture in microwells. In addition, these cell culture methods are simple to use and implement, yet versatile, and have the potential to provide a wide range of microenvironments to improve in vivo-like behavior for a number of cell types and applications. The air bubble masking technique can also be used to produce a micron thick layer of collagen film suspended on top of the microwells. These collagen film enclosed microwells could provide an easy way for high throughput drug screening and cytotoxicity assays as different drug compounds could be pre-loaded and dried in selected microwells and then released during cell culture.
MeSH term(s)	Air ; Cell Culture Techniques/instrumentation ; Hep G2 Cells ; Hepatocytes/cytology ; Humans ; Microfluidics/instrumentation ; Tumor Cells, Cultured
Language	English
Publishing date	2015-02-21
Publishing country	England
Document type	Journal Article
ZDB-ID	2056646-3
ISSN	1473-0189 ; 1473-0197
ISSN (online)	1473-0189
ISSN	1473-0197
DOI	10.1039/c4lc01178c
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Microstructured multi-well plate for three-dimensional packed cell seeding and hepatocyte cell culture.

Goral, Vasiliy N / Au, Sam H / Faris, Ronald A / Yuen, Po Ki

Biomicrofluidics

2014 Volume 8, Issue 4, Page(s) 46502

Abstract: In this article, we present a microstructured multi-well plate for enabling three-dimensional (3D) high density seeding and culture of cells through the use of a standard laboratory centrifuge to promote and maintain 3D tissue-like cellular morphology ... ...

Abstract	In this article, we present a microstructured multi-well plate for enabling three-dimensional (3D) high density seeding and culture of cells through the use of a standard laboratory centrifuge to promote and maintain 3D tissue-like cellular morphology and cell-specific functionality in vitro without the addition of animal derived or synthetic matrices or coagulants. Each well has microfeatures on the bottom that are comprised of a series of ditches/open microchannels. The dimensions of the microchannels promote and maintain 3D tissue-like cellular morphology and cell-specific functionality in vitro. After cell seeding with a standard pipette, the microstructured multi-well plates were centrifuged to tightly pack cells inside the ditches in order to enhance cell-cell interactions and induce formation of 3D cellular structures during cell culture. Cell-cell interactions were optimized based on cell packing by considering dimensions of the ditches/open microchannels, orientation of the microstructured multi-well plate during centrifugation, cell seeding density, and the centrifugal force and time. With the optimized cell packing conditions, we demonstrated that after 7 days of cell culture, primary human hepatocytes adhered tightly together to form cord-like structures that resembled 3D tissue-like cellular architecture. Importantly, cell membrane polarity was restored without the addition of animal derived or synthetic matrices or coagulants.
Language	English
Publishing date	2014-08-15
Publishing country	United States
Document type	Journal Article
ISSN	1932-1058
ISSN	1932-1058
DOI	10.1063/1.4892978
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Evaluation of a Novel Renewable Hepatic Cell Model for Prediction of Clinical CYP3A4 Induction Using a Correlation-Based Relative Induction Score Approach.

Zuo, Rongjun / Li, Feng / Parikh, Sweta / Cao, Li / Cooper, Kirsten L / Hong, Yulong / Liu, Jin / Faris, Ronald A / Li, Daochuan / Wang, Hongbing

Drug metabolism and disposition: the biological fate of chemicals

2017 Volume 45, Issue 2, Page(s) 198–207

Abstract: Metabolism enzyme induction-mediated drug-drug interactions need to be carefully characterized in vitro for drug candidates to predict in vivo safety risk and therapeutic efficiency. Currently, both the Food and Drug Administration and European Medicines ...

Abstract	Metabolism enzyme induction-mediated drug-drug interactions need to be carefully characterized in vitro for drug candidates to predict in vivo safety risk and therapeutic efficiency. Currently, both the Food and Drug Administration and European Medicines Agency recommend using primary human hepatocytes as the gold standard in vitro test system for studying the induction potential of candidate drugs on cytochrome P450 (CYP), CYP3A4, CYP1A2, and CYP2B6. However, primary human hepatocytes are known to bear inherent limitations such as limited supply and large lot-to-lot variations, which result in an experimental burden to qualify new lots. To overcome these shortcomings, a renewable source of human hepatocytes (i.e., Corning HepatoCells) was developed from primary human hepatocytes and was evaluated for in vitro CYP3A4 induction using methods well established by the pharmaceutical industry. HepatoCells have shown mature hepatocyte-like morphology and demonstrated primary hepatocyte-like response to prototypical inducers of all three CYP enzymes with excellent consistency. Importantly, HepatoCells retain a phenobarbital-responsive nuclear translocation of human constitutive androstane receptor from the cytoplasm, characteristic to primary hepatocytes. To validate HepatoCells as a useful tool to predict potential clinical relevant CYP3A4 induction, we tested three different lots of HepatoCells with a group of clinical strong, moderate/weak CYP3A4 inducers, and noninducers. A relative induction score calibration curve-based approach was used for prediction. HepatoCells showed accurate prediction comparable to primary human hepatocytes. Together, these results demonstrate that Corning HepatoCells is a reliable in vitro model for drug-drug interaction studies during the early phase of drug testing.
MeSH term(s)	Cells, Cultured ; Cytochrome P-450 CYP3A/biosynthesis ; Cytochrome P-450 CYP3A/genetics ; Cytochrome P-450 CYP3A Inducers/pharmacology ; Dose-Response Relationship, Drug ; Drug Evaluation, Preclinical ; Female ; Genotype ; Hepatocytes/drug effects ; Hepatocytes/enzymology ; Humans ; Models, Biological ; Predictive Value of Tests ; Primary Cell Culture ; Receptors, Cytoplasmic and Nuclear/genetics ; Receptors, Cytoplasmic and Nuclear/metabolism
Chemical Substances	Cytochrome P-450 CYP3A Inducers ; Receptors, Cytoplasmic and Nuclear ; constitutive androstane receptor (438XLITDI3) ; Cytochrome P-450 CYP3A (EC 1.14.14.1) ; CYP3A4 protein, human (EC 1.14.14.55)
Language	English
Publishing date	2017-01-06
Publishing country	United States
Document type	Journal Article
ZDB-ID	186795-7
ISSN	1521-009X ; 0090-9556
ISSN (online)	1521-009X
ISSN	0090-9556
DOI	10.1124/dmd.116.072124
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Zs.A 1014: Show issues

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Article: Effects of sodium butyrate and acidic fibroblast growth factor on TDC32300 cultured cells.

Pienvichit, Paneeya / Konkin, Tamako A / Faris, Ronald A

Journal of the Medical Association of Thailand = Chotmaihet thangphaet

2002 Volume 85 Suppl 4, Page(s) S1089–95

Abstract: Background: Many studies have demonstrated that transition duct cells (TDC) are facultative liver stem cells. Our laboratory established TDC32300 cell lines with hepatic progenitor markers. The authors proposed that cell culture using sodium butyrate ( ... ...

Abstract	Background: Many studies have demonstrated that transition duct cells (TDC) are facultative liver stem cells. Our laboratory established TDC32300 cell lines with hepatic progenitor markers. The authors proposed that cell culture using sodium butyrate (NaBut) and acidic fibroblast growth factors (aFGF) may support the differentiation of TDC32300 cells along the hepatic lineage. Methods: TDC32300 cells were cultured in four different conditions 1) STON media alone; 2) STON with NaBut in 3 different concentrations, 1 mM, 3.75 mM and 5 mM; 3) STON with aFGF; and 4) STON with aFGF and dexamethasone. After day 5, the cultured cells were fixed and stained with monoclonal antibodies to rat liver antigens and anti-proliferating nuclear antigen (PCNA). Results: Proliferation of TDC32300 cells cultured in the high concentration of NaBut (3.75 and 5 mM) was inhibited. This phenomenon was confirmed by the reduction in cell number and decrease in PCNA expression. Irrespective of the concentration, NaBut did not alter the phenotype of the TDC32300 cultured cells. aFGF with or without dexamethasone also did not alter the phenotypic characteristic of TDC32300 cells. Conclusion: TDC32300 cells may not be the hepatic progenitors or that their differentiation may require other culture conditions.
MeSH term(s)	Butyrates/pharmacology ; Cells, Cultured ; Fibroblast Growth Factor 1/pharmacology ; Hepatocytes/drug effects ; Humans ; In Vitro Techniques ; Stem Cells/drug effects
Chemical Substances	Butyrates ; Fibroblast Growth Factor 1 (104781-85-3)
Language	English
Publishing date	2002-11
Publishing country	Thailand
Document type	Journal Article
ZDB-ID	801630-6
ISSN	0125-2208 ; 0025-7036
ISSN	0125-2208 ; 0025-7036
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Zs.A 957: Show issues

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Article: Perfusion-based microfluidic device for three-dimensional dynamic primary human hepatocyte cell culture in the absence of biological or synthetic matrices or coagulants.

Goral, Vasiliy N / Hsieh, Yi-Cheng / Petzold, Odessa N / Clark, Jeffery S / Yuen, Po Ki / Faris, Ronald A

Lab on a chip

2010 Volume 10, Issue 24, Page(s) 3380–3386

Abstract: We describe a perfusion-based microfluidic device for three-dimensional (3D) dynamic primary human hepatocyte cell culture. The microfluidic device was used to promote and maintain 3D tissue-like cellular morphology and cell-specific functionality of ... ...

Abstract	We describe a perfusion-based microfluidic device for three-dimensional (3D) dynamic primary human hepatocyte cell culture. The microfluidic device was used to promote and maintain 3D tissue-like cellular morphology and cell-specific functionality of primary human hepatocytes by restoring membrane polarity and hepatocyte transport function in vitro without the addition of biological or synthetic matrices or coagulants. A unique feature of our dynamic cell culture device is the creation of a microenvironment, without the addition of biological or synthetic matrices or coagulants, that promotes the 3D organization of hepatocytes into cord-like structures that exhibit functional membrane polarity as evidenced by the expression of gap junctions and the formation of an extended, functionally active, bile canalicular network.
MeSH term(s)	Adenosine Triphosphate/chemistry ; Bile Canaliculi/cytology ; Cell Culture Techniques/instrumentation ; Cell Culture Techniques/methods ; Cells, Cultured ; Coagulants/chemistry ; Equipment Design ; Gap Junctions ; Hepatocytes/cytology ; Humans ; Imaging, Three-Dimensional/methods ; Microfluidic Analytical Techniques ; Microscopy, Fluorescence/methods ; Models, Biological ; Multidrug Resistance-Associated Proteins/metabolism ; Perfusion
Chemical Substances	Coagulants ; Multidrug Resistance-Associated Proteins ; multidrug resistance-associated protein 2 (4AF605U6JN) ; Adenosine Triphosphate (8L70Q75FXE)
Language	English
Publishing date	2010-12-21
Publishing country	England
Document type	Journal Article
ZDB-ID	2056646-3
ISSN	1473-0189 ; 1473-0197
ISSN (online)	1473-0189
ISSN	1473-0197
DOI	10.1039/c0lc00135j
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Article: White matter injury after cerebral ischemia in ovine fetuses.

Petersson, Katherine H / Pinar, Halit / Stopa, Edward G / Faris, Ronald A / Sadowska, Grazyna B / Hanumara, R Choudry / Stonestreet, Barbara S

Pediatric research

2002 Volume 51, Issue 6, Page(s) 768–776

Abstract: The effects of cerebral ischemia on white matter changes in ovine fetuses were examined after exposure to bilateral carotid artery occlusion. Fetal sheep were exposed to 30 min of ischemia followed by 48 (I/R-48, n = 8) or 72 (I/R-72, n = 10) h of ... ...

Abstract	The effects of cerebral ischemia on white matter changes in ovine fetuses were examined after exposure to bilateral carotid artery occlusion. Fetal sheep were exposed to 30 min of ischemia followed by 48 (I/R-48, n = 8) or 72 (I/R-72, n = 10) h of reperfusion or control sham treatment (control, n = 4). Serial coronal sections stained with Luxol fast blue/hematoxylin and eosin were scored for white matter, cerebral cortical, and hippocampal lesions. All areas received graded pathologic scores of 0 to 5, reflecting the degree of injury where 0 = 0%, 1 = 1% to 25%, 2 = 26% to 50%, 3 = 51% to 75%, 4 = 76% to 95%, and 5 = 96% to 100% of the area damaged. Dual-label immunofluorescence using antibodies against glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP) were used to characterize white matter lesions. Basic fibroblast growth factor (FGF-2) was measured in the frontal cortex by ELISA. Results of the pathologic scores showed that the white matter of the I/R-72 (2.74 +/- 0.53, mean +/- SEM) was more (p < 0.05) damaged when compared with the control (0.80 +/- 0.33) group. Cortical lesions were greater (p < 0.05) in the I/R-48 (2.12 +/- 0.35) than the control (0.93 +/- 0.09) group. White matter lesions were characterized by reactive GFAP-positive astrocytes and a loss of MBP in oligodendrocytes. The ratio of MBP to GFAP decreased (p < 0.05) as a function of ischemia, indicative of a proportionally greater loss of MBP than GFAP. FGF-2 concentrations were higher (p < 0.05) in the I/R-72 than the control group and there was a direct correlation between the pathologic scores (PS) and FGF-2 concentrations (FGF-2 = e((1.6 PS-0.90)) + 743, n = 17, r = 0.73, p < 0.001). We conclude that carotid artery occlusion results in quantifiable white matter lesions that are associated with a loss of MBP from myelin, and that FGF-2, a purported mediator of recovery from brain injury in adult subjects, increases in concentration in proportion to the severity of brain damage in the fetus.
MeSH term(s)	Animals ; Astrocytes/chemistry ; Astrocytes/pathology ; Brain Ischemia/pathology ; Carbon Dioxide/blood ; Cerebral Cortex/chemistry ; Cerebral Cortex/embryology ; Cerebral Cortex/pathology ; Coloring Agents ; Eosine Yellowish-(YS) ; Female ; Fetus/chemistry ; Fetus/pathology ; Fibroblast Growth Factor 2/analysis ; Glial Fibrillary Acidic Protein/analysis ; Hematoxylin ; Hippocampus/embryology ; Hippocampus/pathology ; Indoles ; Myelin Basic Protein/analysis ; Nerve Fibers/chemistry ; Nerve Fibers/pathology ; Oligodendroglia/chemistry ; Oligodendroglia/pathology ; Oxygen/blood ; Pregnancy ; Sheep
Chemical Substances	Coloring Agents ; Glial Fibrillary Acidic Protein ; Indoles ; Myelin Basic Protein ; Fibroblast Growth Factor 2 (103107-01-3) ; Luxol Fast Blue MBS (1328-51-4) ; Carbon Dioxide (142M471B3J) ; Oxygen (S88TT14065) ; Eosine Yellowish-(YS) (TDQ283MPCW) ; Hematoxylin (YKM8PY2Z55)
Language	English
Publishing date	2002-06
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	4411-8
ISSN	1530-0447 ; 0031-3998
ISSN (online)	1530-0447
ISSN	0031-3998
DOI	10.1203/00006450-200206000-00019
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Zs.A 564: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 373: Show issues

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Article ; Online: An Antigenic Portrait of the Liver during Carcinogenesis

Hixson, Douglas C. / Faris, Ronald A. / Thompson, Nancym L.

Pathobiology - Exploring the basis of disease

1990 Volume 58, Issue 2, Page(s) 65–77

Language	English
Publisher	S. Karger AG
Publishing place	Basel
Publishing country	Switzerland
Document type	Article ; Online
ZDB-ID	1022703-9
ISSN	1423-0291 ; 1015-2008 ; 1015-2008
ISSN (online)	1423-0291
ISSN	1015-2008
DOI	10.1159/000163565
Database	Karger publisher's database

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Ud II Zs.101: Show issues

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Article ; Online: Delineation of Antigenic Pathways of Ethionine-lnduced Liver Cancer in the Rat

Hixson, Douglas C. / Afflgne, Suzanne / Faris, Ronald A. / McBride, Angela C.

Pathobiology - Exploring the basis of disease

1996 Volume 64, Issue 2, Page(s) 79–90

Abstract: Although oval cell proliferation is observed prior to the appearance of hepatic nodules and hepatocellular carcinomas during ethionine-induced liver carcinogenesis in the rat, the role of these presumptive hepatic stem cells during the neoplastic process ...

Abstract	Although oval cell proliferation is observed prior to the appearance of hepatic nodules and hepatocellular carcinomas during ethionine-induced liver carcinogenesis in the rat, the role of these presumptive hepatic stem cells during the neoplastic process remains controversial. In order to investigate this question, we have used a panel of monoclonal antibodies against antigens associated with normal hepatocytes, oval cells and transplantable hepatocellular carcinomas (THC) to trace antigenic pathways leading to liver cancer. Male ACI rats were fed a choline-deficient diet containing 0.1 % DL-ethionine for 4, 16 or 30 weeks. Immunocytochemical analysis of frozen liver sections revealed a sub-population of hepatic nodules (7/52), carcinomas (8/15) and lung metastases (3/5) containing populations of cells expressing both oval cell, hepatocyte and neoplastic markers. Carcinomas expressing oval cell markers often appeared as a mosaic of well-defined patches composed of phenotypically distinct cells. Many of the phenotypes expressed closely mimicked patterns of expression observed in fetal and neonatal liver. THC derived from primary tumors positive for oval cell antigens (4/5) continued to express these markers. Northern blot analysis and immunocytochemical analysis revealed that 4/5 primary hepatocellular carcinomas (PHC) and THC expressed α-fetoprotein (AFP) and albumin transcripts and contained subpopulations expressing AFP together with hepatocyte and oval antigens. In contrast, a well-differentiated PHC and its corresponding THC lacked AFP mRNA and oval cell antigens but showed strong expression of both hepatocyte and neoplastic markers. These results demonstrate that a subpopulation of malignant and metastatic hepatocellular carcinomas are comprised of cells expressing multiple oval cell markers in this model system.
Keywords	Ethionine ; Oval cells ; Hepatocellular carcinoma ; Monoclonal antibodies
Language	English
Publisher	S. Karger AG
Publishing place	Basel
Publishing country	Switzerland
Document type	Article ; Online
ZDB-ID	1022703-9
ISSN	1423-0291 ; 1015-2008 ; 1015-2008
ISSN (online)	1423-0291
ISSN	1015-2008
DOI	10.1159/000164013
Database	Karger publisher's database

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Article ; Online: Methods for advanced hepatocyte cell culture in microwells utilizing air bubbles.

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Article: Microstructured multi-well plate for three-dimensional packed cell seeding and hepatocyte cell culture.

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Article ; Online: Evaluation of a Novel Renewable Hepatic Cell Model for Prediction of Clinical CYP3A4 Induction Using a Correlation-Based Relative Induction Score Approach.

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Article: Effects of sodium butyrate and acidic fibroblast growth factor on TDC32300 cultured cells.

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Article: Perfusion-based microfluidic device for three-dimensional dynamic primary human hepatocyte cell culture in the absence of biological or synthetic matrices or coagulants.

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Article: White matter injury after cerebral ischemia in ovine fetuses.

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Article ; Online: An Antigenic Portrait of the Liver during Carcinogenesis

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Article ; Online: Delineation of Antigenic Pathways of Ethionine-lnduced Liver Cancer in the Rat

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