Novel Cell Culture Systems: Nano and Microtechnology for Toxicology

Shuler, M. L.; Xu, Hui

doi:10.1002/9780470145890.ch25

Cited by 2 publications

(4 citation statements)

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“…MicroCCA was designed based on human physiological parameters, and was a physical representation of a human body. Flow rate of the medium recirculation was calculated based on fluid residence time in individual organs 4, 5, 7. The external reservoir represents “other tissue” that were not present on the chip.…”

Section: Restults and Discussionmentioning

confidence: 99%

“…The medium is recirculated constantly through the reservoir, tubing, and cell culture chambers, which mimics the blood circulation pattern in the body. Such a system can address issues of reactive metabolite formation and tissue–tissue interaction through exchange of metabolites or signal molecules 4, 7. Other experimental parameters such as the period of experiments were based on experimental protocols for use of a microCCA.…”

Section: Restults and Discussionmentioning

confidence: 99%

“…In addition to microbore tubing, most microfluidic devices use polymers such as Plexiglas, PDMS, polycarbonate and other plastics as body or housing materials. In microCCA devices developed in our lab, the silicon microCCA chip is sandwiched between two Plexiglas or polycarbonate pieces 4, 5, 7. Medium recirculating inside the channels and cell culture chambers is in direct contact with the Plexiglas or polycarbonate housing, which may cause chemical loss or contaminations as well as with the tubing.…”

Section: Restults and Discussionmentioning

confidence: 99%

“…Materials used for microfluidic cell culture devices may be glass, silicon, ceramics, metal, plastics, or polymer hydrogels 5–10. However, disposable plastic materials are most popular because of their low cost and disposable nature, which significantly reduces preparation and cleanup time, avoids cross‐contamination, and makes devices lightweight and portable.…”

Section: Introductionmentioning

confidence: 99%

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Quantification of chemical–polymer surface interactions in microfluidic cell culture devices

Shuler

2009

Biotechnology Progress

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Microfluidic cell culture devices have been used for drug development, chemical analysis, and environmental pollutant detection. Because of the decreased fluid volume and increased surface area to volume ratio, interactions between device surfaces and the fluid is a key element that affects the performance and detection accuracy of microfluidic devices, particularly if fluid is recirculated by a peristaltic pump. However, this issue has not been studied in detail in a microfluidic cell culture environment. In this study, chemical loss and contaminant leakage from various polymer surfaces in a microfluidic setup were characterized. The effects of hydrophilic coating with Poly (vinyl alcohol), Pluronic(R) F-68, and multi-layer ionic coating were measured. We observed significant surface adsorption of estradiol, doxorubicin, and verapamil with PharMed BPT tubing, whereas PTFE/BPT and stainless steel/BPT hybrid tubing caused less chemical loss in proportion to the fraction of BPT tubing in the hybrid system. Contaminants leaching out of the BPT tubing were found to be estrogen receptor agonists as determined by estrogen-induced green fluorescence expression in an estrogen responsive Ishikawa cell line and also caused interference with an estradiol enzyme-linked immunosorbent assay (ELISA) assay. Stainless steel/BPT hybrid tubing caused the least interference with ELISA. In summary, polymer surface and chemical interactions inside microfluidic systems should not be neglected and require careful investigations when results from a microfluidic system are compared with results from a macroscale cell culture setup.

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Section: Restults and Discussionmentioning

confidence: 99%

Section: Restults and Discussionmentioning

confidence: 99%

Section: Restults and Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Quantification of chemical–polymer surface interactions in microfluidic cell culture devices

Shuler

2009

Biotechnology Progress

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Development of a stable dual cell‐line GFP expression system to study estrogenic endocrine disruptors

Kraus

Shuler

2008

Biotech & Bioengineering

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Environmental estrogenic endocrine disruptors are a health concern. Here we constructed a dual cell-line green fluorescence protein (GFP) expression system to identify and study endocrine disrupting compounds with activities of estrogen receptor agonists or antagonists. Human breast cancer MCF-7 cells and endometrial carcinoma Ishikawa cells were infected with a two tandem estrogen response elements--E4 promoter-GFP reporter gene construct. The use of GFP reporter enabled direct and simple evaluations of cell responses. GFP intensity in stably transfected MCF7-GFP and Ishikawa-GFP cells was dose-responsive to 17-beta-estradiol, diethylstilbestrol, 2-hydroxyestradiol, and environmental toxins bisphenol A, genistein and o-p'-DDT. Raloxifene and tamoxifen were effective antiestrogens in MCF7-GFP cells, but acted as partial estrogen receptor agonists in Ishikawa-GFP cells at concentrations of 0.1 nM and above. No synergistic effect was observed in chemical combinations between organochlorine pesticides methoxychlor, o-p'-DDT, p-p'-DDT, nor between estradiol and estrone. In summary, for the first time the effects of estrogen receptor agonists or antagonists were compared between mammary and endometrial cancer cells both stably expressing identical plasmids with GFP reporter genes under the control of tandem estrogen response elements. This dual cell-line system provides a rapid method and sensitive assay to identify environmental estrogens, antiestrogens, selective estrogen receptor modulators and to study their tissue specific effects and chemical interactions. Such a system is especially useful for direct and parallel toxicity assessments with a microfluidic cell culture device.

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Novel Cell Culture Systems: Nano and Microtechnology for Toxicology

Cited by 2 publications

References 0 publications

Quantification of chemical–polymer surface interactions in microfluidic cell culture devices

Quantification of chemical–polymer surface interactions in microfluidic cell culture devices

Development of a stable dual cell‐line GFP expression system to study estrogenic endocrine disruptors

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