Exposure of BALB/3T3 fibroblast cells to temporal concentration profile of toxicant inside microfluidic device

다국어 초록 (Multilingual Abstract)

This work describes a simple method that generates temporal concentration profiles of chemicals inside a microfluidic channel and exposes cultured cells to chemicals in order to mimic the exponential clearance curve of chemicals in the human body. The device fabricated with a poly (dimethylsiloxane)(PDMS) replica mold and a glass substrate was used to create the microfluidic channels. The fluid streams inside the channels were controlled by programmable syringe pumps. By controlling the relative flow rates of the fluidic inlets using separate syringe pumps, the resulting composition of the inlets that feed the concentration generator zone can be controlled to generate temporal profiles of chemicals in the observation zone.
The concentration profiles of fluorescent dye were confirmed by analyzing the time-lapse fluorescence images. Finally, a temporal concentration profile of toxic cadmium solution with exponential clearance curve was exposed to BALB/3T3 fibroblast cells confluently cultured inside a microfluidic device for 3 h, and the behavior of the cells was monitored using a time-lapse microscope. The relative concentration of reactive oxygen species inside the exposed cells was measured indirectly by using a fluorescent probe to assess the degree of cell death. The method developed in this work offers a convenient way of controlling the concentration of chemicals inside microfluidic channels, and enables time-dependent exposure of chemicals to cells and monitoring of the behavior of cells for biological research applications.

참고문헌 (Reference)

1 Poulsen, C.R, "Static and dynamic acute cytotoxicity assays on microfluidic devices" 77 : 667-672, 2005

2 Whitesides, G.M, "Soft lithography in biology and biochemistry" 3 : 335-373, 2001

3 Chien, R.-L, "Multiport flow-control system for lab-on-a-chip microfluidic devices" 371 : 106-111, 2001

4 Zhang, X, "Microfluidic system for generation of sinusoidal glucose waveforms for entrainment of islets of langerhans" 82 : 6704-6711, 2010

5 Zhang, X, "Microfluidic perfusion system for automated delivery of temporal gradients to islets of langerhans" 81 : 1162-1168, 2009

6 Taylor, A.M, "Microfluidic multicompartment device for neuroscience research" 19 : 1551-1556, 2003

7 Cao, L, "Microfluidic multi-analyte gradient generator" 398 : 1985-1991, 2010

8 Pihl, J, "Microfluidic gradient-generating device for pharmacological profiling" 77 : 3897-3903, 2005

9 Hung, L.-H, "Microfluidic devices for the synthesis of nanoparticles and biomaterials" 27 : 1-6, 2007

10 Bennett, M.R, "Microfluidic devices for measuring gene network dynamics in single cells" 10 : 628-638, 2009

11 Marle, L, "Microfluidic devices for environmental monitoring" 24 : 795-802, 2005

12 Andersson, H, "Microfluidic devices for cellomics: a review" 92 : 315-325, 2003

13 Cimetta, E, "Microfluidic device generating stable concentration gradients for long term cell culture: application to Wnt3a regulation of β-catenin signaling" 10 : 3277-3283, 2010

14 Yeo, L.Y, "J.R. Microfluidic devices for bioapplications" 7 : 12-48, 2011

15 Wang, Z, "High-density microfluidic arrays for cell cytotoxicity analysis" 7 : 740-745, 2007

16 Lin, F, "Generation of dynamic temporal and spatial concentration gradients using microfluidic devices" 4 : 164-167, 2004

17 Azizi, F, "Generation of dynamic chemical signals with pulse code modulators" 8 : 907-912, 2008

18 Kim, M, "Diffusion-based and long-range concentration gradients of multiple chemicals for bacterial chemotaxis assays" 82 : 9401-9409, 2010

19 Evenhuis, C.J, "Determination of inorganic ions using microfluidic devices" 25 : 3602-3624, 2004

20 Sarrazin, F, "Chemical reaction imaging within microfluidic devices using confocal raman spectroscopy: The case of water and deuterium oxide as a model system" 80 : 1689-1695, 2008

21 Fair, R.B, "Chemical and biological applications of digital-microfluidic devices" 24 : 10-24, 2007

22 Mahto, S.K, "Assessment of cytocompatibility of surface-modified CdSe/ZnSe quantum dots for BALB/3T3 fibroblast cells" 24 : 1070-1077, 2010

23 Skurtys, O, "Applications of microfluidic devices in food engineering" 3 : 1-15, 2008

24 Li, J, "Application of microfluidic devices to proteomics research" 1 : 157-168, 2002

25 Diao, J, "A three-channel microfluidic device for generating static linear gradients and its application to the quantitative analysis of bacterial chemotaxis" 6 : 381-388, 2006

26 Dai, W, "A prototypic microfluidic platform generating stepwise concentration gradients for real-time study of cell apoptosis" 4 : 024101-, 2010

27 Olofsson, J, "A microfluidics approach to the problem of creating separate solution environments accessible from macroscopic volumes" 76 : 4968-4976, 2004

28 Ainla, A, "A microfluidic diluter based on pulse width flow modulation" 81 : 5549-5556, 2009

29 Walker, G.M, "A linear dilution microfluidic device for cytotoxicity assays" 7 : 226-232, 2007