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Article ; Online: Microfluidic platforms for hepatocyte cell culture: new technologies and applications.

Goral, Vasiliy N / Yuen, Po Ki

2012 Volume 40, Issue 6, Page(s) 1244–1254

Abstract: In this article, we summarize the key elements of microfluidic platforms for mimicking in vivo hepatocyte cell culture and the major recent advances in this area. Specifically, we will give brief background and rationale for key design requirements for ... ...

Abstract	In this article, we summarize the key elements of microfluidic platforms for mimicking in vivo hepatocyte cell culture and the major recent advances in this area. Specifically, we will give brief background and rationale for key design requirements for mimicking in vivo hepatocyte cell culture, and then summarize findings, applications, and limitations from microfluidic platforms that addressed these design requirements. Although no ideal microfluidic platform has so far been developed for fully mimicking in vivo hepatocyte cell culture, some approaches and designs have demonstrated great potential in this area.
MeSH term(s)	Animals ; Cell Culture Techniques/instrumentation ; Cell Culture Techniques/methods ; Hepatocytes/cytology ; Hepatocytes/metabolism ; Humans ; Microfluidic Analytical Techniques/instrumentation ; Microfluidic Analytical Techniques/methods
Language	English
Publishing date	2012-06
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	185984-5
ISSN	1573-9686 ; 0191-5649 ; 0090-6964
ISSN (online)	1573-9686
ISSN	0191-5649 ; 0090-6964
DOI	10.1007/s10439-011-0453-8
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: A pump-free membrane-controlled perfusion microfluidic platform.

Goral, Vasiliy N / Tran, Elizabeth / Yuen, Po Ki

Biomicrofluidics

2015 Volume 9, Issue 5, Page(s) 54103

Abstract: In this article, we present a microfluidic platform for passive fluid pumping for pump-free perfusion cell culture, cell-based assay, and chemical applications. By adapting the passive membrane-controlled pumping principle from the previously developed ... ...

Abstract	In this article, we present a microfluidic platform for passive fluid pumping for pump-free perfusion cell culture, cell-based assay, and chemical applications. By adapting the passive membrane-controlled pumping principle from the previously developed perfusion microplate, which utilizes a combination of hydrostatic pressure generated by different liquid levels in the wells and fluid wicking through narrow strips of a porous membrane connecting the wells to generate fluid flow, a series of pump-free membrane-controlled perfusion microfluidic devices was developed and their use for pump-free perfusion cell culture and cell-based assays was demonstrated. Each pump-free membrane-controlled perfusion microfluidic device comprises at least three basic components: an open well for generating fluid flow, a micron-sized deep chamber/channel for cell culture or for fluid connection, and a wettable porous membrane for controlling the fluid flow. Each component is fluidically connected either by the porous membrane or by the micron-sized deep chamber/channel. By adapting and incorporating the passive membrane-controlled pumping principle into microfluidic devices, all the benefits of microfluidic technologies, such as small sample volumes, fast and efficient fluid exchanges, and fluid properties at the micro-scale, can be fully taken advantage of with this pump-free membrane-controlled perfusion microfluidic platform.
Language	English
Publishing date	2015-09-02
Publishing country	United States
Document type	Journal Article
ISSN	1932-1058
ISSN	1932-1058
DOI	10.1063/1.4930120
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Low-cost rapid prototyping of flexible microfluidic devices using a desktop digital craft cutter.

Yuen, Po Ki / Goral, Vasiliy N

Lab on a chip

2010 Volume 10, Issue 3, Page(s) 384–387

Abstract: Low-cost and straight forward rapid prototyping of flexible microfluidic devices using a desktop digital craft cutter is presented. This rapid prototyping method can consistently achieve microchannels as thin as 200 microm in width and can be used to ... ...

Abstract	Low-cost and straight forward rapid prototyping of flexible microfluidic devices using a desktop digital craft cutter is presented. This rapid prototyping method can consistently achieve microchannels as thin as 200 microm in width and can be used to fabricate three-dimensional (3D) microfluidic devices using only double-sided pressure sensitive adhesive (PSA) tape and laser printer transparency film. Various functional microfluidic devices are demonstrated with this rapid prototyping method. The complete fabrication process from device design concept to working device can be completed in minutes without the need of expensive equipment.
Language	English
Publishing date	2010-02-07
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/b918089c
Database	MEDical Literature Analysis and Retrieval System OnLINE

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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

Lab on a chip

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: A continuous perfusion microplate for cell culture.

Goral, Vasiliy N / Zhou, Chunfeng / Lai, Fang / Yuen, Po Ki

Lab on a chip

2013 Volume 13, Issue 6, Page(s) 1039–1043

Abstract: We describe a 96-well microplate with fluidically connected wells that enables the continuous fluid perfusion between wells without the need for external pumping. A single unit in such a perfusion microplate consists of three wells: a source well, a ... ...

Abstract	We describe a 96-well microplate with fluidically connected wells that enables the continuous fluid perfusion between wells without the need for external pumping. A single unit in such a perfusion microplate consists of three wells: a source well, a sample (cell culture) well in the middle and a waste well. Fluid perfusion is achieved using a combination of the hydrostatic pressure generated by different liquid levels in the wells and the fluid wicking through narrow strips of a cellulose membrane connecting the wells. There is an excellent correspondence between the observed perfusion flow dynamics and the flow simulations based on Darcy's Law. Hepatocytes (C3A cells) cultured for 4 days in the perfusion microplate with no media exchange in the cell culture well had the same viability as hepatocytes exposed to a daily exchange of media. EOC 20 cells that require media conditioned by LADMAC cells were shown to be equally viable in the adjacent cell culture well of the perfusion microplate with LADMAC cells cultured in the source well. Tegafur, a prodrug, when added to primary human hepatocytes in the source well, was metabolized into a cytotoxic metabolite that kills colon cancer cells (HCT 116) cultured in the adjacent cell culture well; no toxicity was observed when only medium was in the source well. These results suggest that the perfusion microplate is a useful tool for a variety of cell culture applications with benefits ranging from labor savings to enabling in vivo-like toxicity studies.
MeSH term(s)	Animals ; Cell Culture Techniques/instrumentation ; Cell Culture Techniques/methods ; Cell Survival/drug effects ; Cells, Cultured ; HCT116 Cells ; Humans ; Mice ; Prodrugs/toxicity
Chemical Substances	Prodrugs
Language	English
Publishing date	2013-03-21
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/c2lc41102d
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: A polystyrene-based microfluidic device with three-dimensional interconnected microporous walls for perfusion cell culture.

Chan, Chung Yu / Goral, Vasiliy N / DeRosa, Michael E / Huang, Tony Jun / Yuen, Po Ki

Biomicrofluidics

2014 Volume 8, Issue 4, Page(s) 46505

Abstract: In this article, we present a simple, rapid prototyped polystyrene-based microfluidic device with three-dimensional (3D) interconnected microporous walls for long term perfusion cell culture. Patterned 3D interconnected microporous structures were ... ...

Abstract	In this article, we present a simple, rapid prototyped polystyrene-based microfluidic device with three-dimensional (3D) interconnected microporous walls for long term perfusion cell culture. Patterned 3D interconnected microporous structures were created by a chemical treatment together with a protective mask and the native hydrophobic nature of the microporous structures were selectively made hydrophilic using oxygen plasma treatment together with a protective mask. Using this polystyrene-based cell culture microfluidic device, we successfully demonstrated the support of four days perfusion cell culture of hepatocytes (C3A cells).
Language	English
Publishing date	2014-08-27
Publishing country	United States
Document type	Journal Article
ISSN	1932-1058
ISSN	1932-1058
DOI	10.1063/1.4894409
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Three-dimensional interconnected microporous poly(dimethylsiloxane) microfluidic devices.

Yuen, Po Ki / Su, Hui / Goral, Vasiliy N / Fink, Katherine A

Lab on a chip

2011 Volume 11, Issue 8, Page(s) 1541–1544

Abstract: This technical note presents a fabrication method and applications of three-dimensional (3D) interconnected microporous poly(dimethylsiloxane) (PDMS) microfluidic devices. Based on soft lithography, the microporous PDMS microfluidic devices were ... ...

Abstract	This technical note presents a fabrication method and applications of three-dimensional (3D) interconnected microporous poly(dimethylsiloxane) (PDMS) microfluidic devices. Based on soft lithography, the microporous PDMS microfluidic devices were fabricated by molding a mixture of PDMS pre-polymer and sugar particles in a microstructured mold. After curing and demolding, the sugar particles were dissolved and washed away from the microstructured PDMS replica revealing 3D interconnected microporous structures. Other than introducing microporous structures into the PDMS replica, different sizes of sugar particles can be used to alter the surface wettability of the microporous PDMS replica. Oxygen plasma assisted bonding was used to enclose the microstructured microporous PDMS replica using a non-porous PDMS with inlet and outlet holes. A gas absorption reaction using carbon dioxide (CO(2)) gas acidified water was used to demonstrate the advantages and potential applications of the microporous PDMS microfluidic devices. We demonstrated that the acidification rate in the microporous PDMS microfluidic device was approximately 10 times faster than the non-porous PDMS microfluidic device under similar experimental conditions. The microporous PDMS microfluidic devices can also be used in cell culture applications where gas perfusion can improve cell survival and functions.
MeSH term(s)	Absorption ; Carbon Dioxide/chemistry ; Dimethylpolysiloxanes/chemistry ; Microfluidic Analytical Techniques/methods ; Porosity ; Water/chemistry
Chemical Substances	Dimethylpolysiloxanes ; Water (059QF0KO0R) ; Carbon Dioxide (142M471B3J) ; baysilon (63148-62-9)
Language	English
Publishing date	2011-04-21
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/c0lc00660b
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Electrochemical microfluidic biosensor for the detection of nucleic acid sequences.

Goral, Vasiliy N / Zaytseva, Natalya V / Baeumner, Antje J

Lab on a chip

2006 Volume 6, Issue 3, Page(s) 414–421

Abstract: A microfluidic biosensor with electrochemical detection for the quantification of nucleic acid sequences was developed. In contrast to most microbiosensors that are based on fluorescence for signal generation, it takes advantage of the simplicity and ... ...

Abstract	A microfluidic biosensor with electrochemical detection for the quantification of nucleic acid sequences was developed. In contrast to most microbiosensors that are based on fluorescence for signal generation, it takes advantage of the simplicity and high sensitivity provided by an amperometric and coulorimetric detection system. An interdigitated ultramicroelectrode array (IDUA) was fabricated in a glass chip and integrated directly with microchannels made of poly(dimethylsiloxane) (PDMS). The assembly was packaged into a Plexiglas housing providing fluid and electrical connections. IDUAs were characterized amperometrically and using cyclic voltammetry with respect to static and dynamic responses for the presence of a reversible redox couple-potassium hexacyanoferrate (ii)/hexacyanoferrate (iii) (ferri/ferrocyanide). A combined concentration of 0.5 microM of ferro/ferricyanide was determined as lower limit of detection with a dynamic range of 5 orders of magnitude. Background signals were negligible and the IDUA responded in a highly reversible manner to the injection of various volumes and various concentrations of the electrochemical marker. For the detection of nucleic acid sequences, liposomes entrapping the electrochemical marker were tagged with a DNA probe, and superparamagnetic beads were coated with a second DNA probe. A single stranded DNA target sequence hybridized with both probes. The sandwich was captured in the microfluidic channel just upstream of the IDUA via a magnet located in the outside housing. Liposomes were lysed using a detergent and the amount of released ferro/ferricyanide was quantified while passing by the IDUA. Optimal location of the magnet with respect to the IDUA was investigated, the effect of dextran sulfate on the hybridization reaction was studied and the amount of magnetic beads used in the assay was optimized. A dose response curve using varying concentrations of target DNA molecules was carried out demonstrating a limit of detection at 1 fmol assay(-1) and a dynamic range between 1 and 50 fmol. The overall assay took 6 min to complete, plus 15-20 min of pre-incubation and required only a simple potentiostat for signal recording and interpretation.
MeSH term(s)	Biosensing Techniques/instrumentation ; Biosensing Techniques/methods ; Dextran Sulfate/chemistry ; Dimethylpolysiloxanes/chemistry ; Electrochemistry ; Electrodes ; Equipment Design ; Microfluidic Analytical Techniques/instrumentation ; Microfluidic Analytical Techniques/methods ; Oxidation-Reduction ; Sensitivity and Specificity ; Sequence Analysis, DNA/instrumentation ; Sequence Analysis, DNA/methods ; Silicones/chemistry ; Time Factors
Chemical Substances	Dimethylpolysiloxanes ; Silicones ; baysilon (63148-62-9) ; Dextran Sulfate (9042-14-2)
Language	English
Publishing date	2006-03
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2056646-3
ISSN	1473-0189 ; 1473-0197
ISSN (online)	1473-0189
ISSN	1473-0197
DOI	10.1039/b513239h
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Development of a microfluidic biosensor module for pathogen detection.

Zaytseva, Natalya V / Goral, Vasiliy N / Montagna, Richard A / Baeumner, Antje J

Lab on a chip

2005 Volume 5, Issue 8, Page(s) 805–811

Abstract: The development of a microfluidic biosensor module with fluorescence detection for the identification of pathogenic organisms and viruses is presented in this article. The microfluidic biosensor consists of a network of microchannels fabricated in ... ...

Abstract	The development of a microfluidic biosensor module with fluorescence detection for the identification of pathogenic organisms and viruses is presented in this article. The microfluidic biosensor consists of a network of microchannels fabricated in polydimethylsiloxane (PDMS) substrate. The microchannels are sealed with a glass substrate and packed in a Plexiglas housing to provide connection to the macro-world and ensure leakage-free flow operation. Reversible sealing permits easy disassembly for cleaning and replacing the microfluidic channels. The fluidic flow is generated by an applied positive pressure gradient, and the module can be operated under continuous solution flow of up to 80 microL min(-1). The biosensor recognition principle is based on DNA/RNA hybridization and liposome signal amplification. Superparamagnetic beads are incorporated into the system as a mobile solid support and are an essential part of the analysis scheme. In this study, the design, fabrication and the optimization of concentrations and amounts of the different biosensor components are carried out. The total time required for an assay is only 15 min including sample incubation time. The biosensor module is designed so that it can be easily integrated with a micro total analysis system, which will combine sample preparation and detection steps onto a single chip.
MeSH term(s)	Bacteria/isolation & purification ; Biosensing Techniques/instrumentation ; Biosensing Techniques/methods ; DNA Probes ; Fluorescence ; Liposomes ; Microfluidic Analytical Techniques/instrumentation ; Microfluidic Analytical Techniques/methods ; RNA, Bacterial/analysis ; RNA, Messenger/analysis ; RNA, Ribosomal/analysis ; RNA, Viral/analysis ; Viruses/isolation & purification
Chemical Substances	DNA Probes ; Liposomes ; RNA, Bacterial ; RNA, Messenger ; RNA, Ribosomal ; RNA, Viral
Language	English
Publishing date	2005-08
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2056646-3
ISSN	1473-0189 ; 1473-0197
ISSN (online)	1473-0189
ISSN	1473-0197
DOI	10.1039/b503856a
Database	MEDical Literature Analysis and Retrieval System OnLINE

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