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Design of parallel microfluidic gradient-generating
networks for studying cellular response to chemical stimuli |
| WANG Lihui1, WANG Bo1, SUN Jie1, LI Lianhong1, LIU Dayu2 |
| 1.Department of Pathology, Dalian Medical University; 2.Dalian Institute of Chemical Physics, Chinese Academy of Sciences；Biomedical Engineering Center, Improve Medical Instrument's Co., Ltd.; |
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Abstract A microfluidic chip featuring laminar flow-based parallel gradient-generating networks was designed and fabricated. The microchip contains 5 gradient generators and 30 cell chambers where the resulting concentration gradients of drugs are delivered to stimulate on-chip cultured cells. The microfluidics exploits the advantage of lab-on-a-chip technology by integrating the generation of drug concentration gradients and a series of cell operations including seeding, culture, stimulation and staining into a chip. The microfluidic network was patterned on a glass wafer, which was further bonded to a PDMS film. A series of weir structures were fabricated on the cell culture reservoir to facilitate cell positioning and seeding. Cell injection and fluid delivery were controlled by a syringe pump. Steady parallel concentration gradients were generated by flowing two fluids in each network. Over time observation shows that the microchip was suitable for cell seeding and culture. The microchip described above was applied in studying the role of reduced glutathione (GSH) in mediating chemotherapy sensitivity of MCF-7 cells. MCF-7 cells were treated with concentration gradients of As2O3 and N-acetyl cysteine (NAC) for GSH modulation, followed by exposure to adriamycin. GSH levels were down-regulated upon As2O3 treatment and up-regulated upon NAC treatment. Suppression of intracellular GSH by treatment with As2O3 has been shown to increase sensitivity to adriamycin. Conversely, elevation of intracellular GSH by treatment with NAC leads to increased drug resistance. The integrated microfluidic chip is able to perform multiparametric pharmacological profiling with easy operation, and thus holds great potential for extrapolation to the cell based high-content drug screening.
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Issue Date: 05 December 2008
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| 1 |
El Ali J, Sorger P K, Jensen K F . Cells on chips. Nature, 2006, 442(7101): 403–411.
doi:10.1038/nature05063
|
| 2 |
Chen X, Cui D F, Liu C C, Cai H Y . Chinese JAnal Chem, Microfluidic biochip for bloodcell lysis, 2006 34(11): 1656–1660 (in Chinese)
|
| 3 |
Wu J G, Yue R F, Zeng X F, Kang M, Hu H, Wang Z Y, Liu L T . Studies on digital microfluidic chipfor Micro-Total-Analysis systems. ChineseJ Anal Chem, 2006, 34(7): 1042–1046 (in Chinese)
|
| 4 |
Weigl B H, Bardell R L, Cabrera C R . Lab-on-a-chip for drug development. Adv Drug Deliv Rev, 2003, 55(3): 349–377.
doi:10.1016/S0169-409X(02)00223-5
|
| 5 |
Jeon N L, Dertinger S K, Chiu D T, Choi I S, Stroock A D, Whitesides G M . Generation of solution and surface gradients using microfluidic systems. Langmuir, 2000, 16(22): 8311–8316.
doi:10.1021/la000600b
|
| 6 |
Dertinger S K, Chiu D T, Jeon N L, Whitesides G M . Generation of gradients having complex shapes using microfluidicnetworks. Anal. Chem, 2001, 73(6): 1240–1246.
doi:10.1021/ac001132d
|
| 7 |
Jeon N L, Baskaran H, Dertinger S K, Whitesides G M, van de Water L, Toner M . Neutrophil chemotaxis in linear and complex gradientsof interleukin-8 formed in a microfabricated device. Nat Biotechnol, 2002, 20(8): 826–830
|
| 8 |
Saadi W, Wang S J, Lin F, Jeon N L . Biomed. Aparallel-gradient microfluidic chamber for quantitative analysis ofbreast cancer cell chemotaxis. Microdevices, 2006, 8(2): 109–118.
doi:10.1007/s10544-006-7706-6
|
| 9 |
Thompson D M, King K R, Wieder K J, Toner M, Yarmush M L, Jayaraman A . Dynamicgene expression profiling using a microfabricated living cell array. Anal Chem, 2004, 76(14): 4098–4103.
doi:10.1021/ac0354241
|
| 10 |
Liu D Y, Shi M, Huang H, Long Z, Zhou X, Qin J, Lin B C . Isotachophoresis preconcentrationintegrated microfluidic chip for highly sensitive genotyping of thehepatitis B virus J. Chromatogr B, 2006, 21(844): 32–38
|
| 11 |
Vahter M, Concha G . Pharmacol. Role of metabolismin arsenic toxicity. Toxicol, 2001, 89(1): 1–5
|
| 12 |
Shimizu M, Hochadel J F, Fulmer B A, Waalkes M P . Effect of glutathione depletion and metallothionein gene expressionon arsenic-induced cytotoxicity and c-myc expression in vitro. ToxicolSci, 1998, 45(2): 204–211
|
| 13 |
Yim C Y, Hibbs J B, Mcgregor J R, Galinsky R E, Samlowski W E . Use of N-acetyl cysteineto increase intracellular glutathione during the induction of antitumorresponses by IL-2. J Immunology, 1994, 152(12): 5796–5805
|
| 14 |
Davis W J, Ronai Z, Tew K D . Cellular thiols and reactive oxygen species in drug-inducedapoptosis. J Pharmacol Exp Ther, 2001, 296(1): 1–6
|
| 15 |
Schroder C P, Godwin A K, O'Dwyer P J, Tew K D, Hamilton T C, Ozols R F . Glutathione and drug resistance. CancerInvest, 1996, 14(2): 158–168.
doi:10.3109/07357909609018891
|
| 16 |
Anderson M E . Glutathione: an overview of biosynthesis and modulation. Chem Biol Interact, 1998, 111/112: 1–14.
doi:10.1016/S0009-2797(97)00146-4
|
| 17 |
Ishikawa T, Ali Osman F . Glutathione-associated cis-diamminedichloroplatinum(II)metabolism and ATP-dependent efflux from leukemia cells. Molecularcharacterization of glutathione-platinum complex and its biologicalsignificance. J Biol Chem, 1993, 268(27): 20116–20125
|
| 18 |
Shiga H, Heath E I, Rasmussen A A, Trock B, Johnston P G, Forastiere A A, Langmacher M, Baylor A, Lee M, Cullen K . J. Prognostic value of p53, glutathioneS-transferase pi, and thymidylate synthase for neoadjuvant cisplatin-basedchemotherapy in head and neck cancer. ClinCancer Res, 1999, 5(12): 4097–4104
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