A microfluidic device mimicking acinar concentration gradients across the liver acinus

Springer Science and Business Media LLC - Tập 15 - Trang 767-780 - 2013
Ming-Cheng Shih1,2, Shih-Heng Tseng2,3, Yu-Shih Weng2, I-Ming Chu3, Cheng-Hsien Liu4
1Institute of NanoEngineering and MicroSystem, National Tsing Hua University, Hsinchu, Republic of China
2Biomedical Technology and Device Research Laboratories, Industrial Technology Research Institute, Chutung, Hsinchu 310, Republic of China
3Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Republic of China
4Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Republic of China

Tóm tắt

The acinus-mimicking microfluidic chip, which simulates the in vivo condition of the liver, was developed and reported in this paper. The gradient microenvironment of the liver acinus is replicated within this proposed microfluidic chip. The advantage of this acinus-mimicking chip is capable of adjusting the concentration gradient in a relatively short period of time at around 10 s. At the same instance the non-linear concentration gradient can be presented in the various zones within this microfluidic chip. The other advantage of this proposed design is in the convenience of allowing the direct injection of the cells into the chip. The environment within the chip is multi-welled and gel-free with high cell density. The multi-row pillar microstructure located at the entrance of the top and bottom flow channels is designed to be able to balance the pressure of the perfusion medium. Through this mechanism the shear stress experienced by the cultured cells can be minimized to reduce the potential damage flow from the perfusion process. (3)The fluorescence staining and the observations of the cell morphology verify the life and death of the cells. The shear stress experienced by the cells in the various zones within the chip can be effectively mapped. The serum glutamic oxaloacetic transaminase (SGOT) collected from the supernatants was used to determine the effects of the degassing process and the shear stress of the medium flow on the cultured cells.

Tài liệu tham khảo

V.V. Abhyankar, D.J. Beebe, Spatiotemporal micropatterning of cells on arbitrary substrates. Anal. Chem. 79(11), 4066–4073 (2007) V.V. Abhyankar, M.W. Toepke, C.L. Cortesio et al., A platform for assessing chemotactic migration within a spatiotemporally defined 3D microenvironment. Lab Chip 8(9), 1507–1515 (2008) J.W. Allen, S.N. Bhatia, Formation of steady-state oxygen gradients in vitro: application to liver zonation. Biotechnol. Bioeng. 82(3), 253–262 (2003) J.W. Allen, S.R. Khetani, S.N. Bhatia, In vitro zonation and toxicity in a hepatocyte bioreactor. Toxicol. Sci. 84(1), 110–119 (2005) J. Atencia, J. Morrow, L.E. Locascio, The microfluidic palette: a diffusive gradient generator with spatio-temporal control. Lab Chip 9(18), 2707–2714 (2009) J. Atencia, G.A. Cooksey, L.E. Locascio, A robust diffusion-based gradient generator for dynamic cell assays. Lab Chip 12(2), 309–316 (2012) S.N. Bhatia, U.J. Balis, M.L. Yarmush et al., Effect of cell-cell interactions in preservation of cellular phenotype: cocultivation of hepatocytes and nonparenchymal cells. FASEB J. 13(14), 1883–1900 (1999) T.A. Broughan, R. Naukam, C. Tan et al., Effects of hepatic zonal oxygen levels on hepatocyte stress responses. J. Surg. Res. 145(1), 150–160 (2008) K. Campbell, A. Groisman, Generation of complex concentration profiles in microchannels in a logarithmically small number of steps. Lab Chip 7(2), 264–272 (2007) A. Carraro, W.M. Hsu, K.M. Kulig et al., In vitro analysis of a hepatic device with intrinsic microvascular-based channels. Biomed. Microdevices 10(6), 795–805 (2008) Y.A. Chen, A.D. King, H.C. Shih et al., Generation of oxygen gradients in microfluidic devices for cell culture using spatially confined chemical reactions. Lab Chip. 11(21), 3626–33 (2011) B.G. Chung, J. Choo, Microfluidic gradient platforms for controlling cellular behavior. Electrophoresis. 31(18), 3014–27 (2010) B.G. Chung, L.A. Flanagan, S.W. Rhee et al., Human neural stem cell growth and differentiation in a gradient-generating microfluidic device. Lab Chip 5(4), 401–406 (2005) B.G. Chung, L.F. Kang, A. Khademhosseini, Micro- and nanoscale technologies for tissue engineering and drug discovery applications. Expert Opin. Drug Discov. 2(12), 1653–1668 (2007) A. Dash, W. Inman, K. Hoffmaster et al., Liver tissue engineering in the evaluation of drug safety. Expert Opin. Drug Metab. Toxicol. 5(10), 1159–1174 (2009) N.T. Elliott, F. Yuan, A review of three-dimensional in vitro tissue models for drug discovery and transport studies. J. Pharm. Sci. 100(1), 59–74 (2011) V.N. Goral, and P.K. Yuen, “Microfluidic Platforms for Hepatocyte Cell Culture: New Technologies and Applications,” Ann. Biomed. Eng (2012) L.G. Griffith, G. Naughton, Tissue engineering–current challenges and expanding opportunities. Science 295(5557), 1009–1014 (2002) L.G. Griffith, M.A. Swartz, Capturing complex 3D tissue physiology in vitro. Nat. Rev. Mol. Cell Biol. 7(3), 211–224 (2006) D. Irimia, S.Y. Liu, W.G. Tharp et al., Microfluidic system for measuring neutrophil migratory responses to fast switches of chemical gradients. Lab Chip 6(2), 191–198 (2006) M. Jain, A. Yeung, K. Nandakumar, Induced charge electro-osmotic concentration gradient generator. Biomicrofluidics. 4(1), 14110 (2010) K. Jungermann, N. Katz, Functional specialization of different hepatocyte populations. Physiol. Rev. 69(3), 708–764 (1989) K. Jungermann, T. Kietzmann, Oxygen: modulator of metabolic zonation and disease of the liver. Hepatology 31(2), 255–260 (2000) T.M. Keenan, A. Folch, Biomolecular gradients in cell culture systems. Lab Chip 8(1), 34–57 (2008) T.M. Keenan, C.W. Frevert, A. Wu et al., A new method for studying gradient-induced neutrophil desensitization based on an open microfluidic chamber. Lab Chip. 10(1), 116–22 (2010) M.S. Kim, J.H. Yeon, J.K. Park, A microfluidic platform for 3-dimensional cell culture and cell-based assays. Biomed. Microdevices 9(1), 25–34 (2007) E. Leclerc, K. El Kirat, L. Griscom, In situ micropatterning technique by cell crushing for co-cultures inside microfluidic biochips. Biomed. Microdevices 10(2), 169–177 (2008) E.L. LeCluyse, P.L. Bullock, A. Parkinson et al., Cultured rat hepatocytes. Pharm. Biotechnol. 8, 121–159 (1996) E.L. LeCluyse, R.P. Witek, M.E. Andersen et al., Organotypic liver culture models: meeting current challenges in toxicity testing. Crit. Rev. Toxicol. 42(6), 501–48 (2012) P.J. Lee, P.J. Hung, L.P. Lee, An artificial liver sinusoid with a microfluidic endothelial-like barrier for primary hepatocyte culture. Biotechnol. Bioeng. 97(5), 1340–1346 (2007) F. Lin, C.M. Nguyen, S.J. Wang et al., Neutrophil migration in opposing chemoattractant gradients using microfluidic chemotaxis devices. Ann. Biomed. Eng. 33(4), 475–482 (2005) K.O. Lindros, Zonation of cytochrome P450 expression, drug metabolism and toxicity in liver. Gen. Pharmacol. 28(2), 191–196 (1997) Y. Ling, J. Rubin, Y. Deng et al., A cell-laden microfluidic hydrogel. Lab Chip 7(6), 756–762 (2007) M-C. S. Y.-S. W. T.-C. S. R.-J. G. C.-H. Liu, “A Novel Micro-Tissue Reactor for Maintaining Hepatocytes in Vitro,” MEMS 2006 Conference, pp. 458–461 (2006) R.S. McCuskey, Sinusoidal endothelial cells as an early target for hepatic toxicants. Clin. Hemorheol. Microcirc. 34(1–2), 5–10 (2006) G. Mehta, C.M. Williams, L. Alvarez et al., Synergistic effects of tethered growth factors and adhesion ligands on DNA synthesis and function of primary hepatocytes cultured on soft synthetic hydrogels. Biomaterials. 31(17), 4657–71 (2010) L.J. Millet, M.E. Stewart, R.G. Nuzzo et al., Guiding neuron development with planar surface gradients of substrate cues deposited using microfluidic devices. Lab Chip 10(12), 1525–35 (2010) Y. Nakao, H. Kimura, Y. Sakai et al., Bile canaliculi formation by aligning rat primary hepatocytes in a microfluidic device. Biomicrofluidics. 5(2), 22212 (2011) J. Park, F. Berthiaume, M. Toner et al., Microfabricated grooved substrates as platforms for bioartificial liver reactors. Biotechnol. Bioeng. 90(5), 632–644 (2005) J.Y. Park, C.M. Hwang, S.H. Lee, Gradient generation by an osmotic pump and the behavior of human mesenchymal stem cells under the fetal bovine serum concentration gradient. Lab Chip 7(12), 1673–1680 (2007) J.Y. Park, S.K. Kim, D.H. Woo et al., Differentiation of neural progenitor cells in a microfluidic chip-generated cytokine gradient. Stem Cells 27(11), 2646–2654 (2009) O. Pelkonen, M. Turpeinen, J. Hakkola et al., Inhibition and induction of human cytochrome P450 enzymes: current status. Arch. Toxicol. 82(10), 667–715 (2008) J. Pihl, J. Sinclair, E. Sahlin et al., Microfluidic gradient-generating device for pharmacological profiling. Anal. Chem. 77(13), 3897–3903 (2005) S.R. Quake, A. Scherer, From micro- to nanofabrication with soft materials. Science 290(5496), 1536–1540 (2000) Y. Tanaka, M. Yamato, T. Okano et al., Evaluation of effects of shear stress on hepatocytes by a microchip-based system. Meas. Sci. Technol. 17(12), 3167–3170 (2006) Y.C. Toh, T.C. Lim, D. Tai et al., A microfluidic 3D hepatocyte chip for drug toxicity testing. Lab Chip 9(14), 2026–2035 (2009) Y.C. Toh, K. Blagovic, J. Voldman, Advancing stem cell research with microtechnologies: opportunities and challenges. Integr. Biol. (Camb). 2(7–8), 305–25 (2010) M.A. Unger, H.P. Chou, T. Thorsen et al., Monolithic microfabricated valves and pumps by multilayer soft lithography. Science 288(5463), 113–116 (2000) P.M. van Midwoud, E. Verpoorte, G.M. Groothuis, Microfluidic devices for in vitro studies on liver drug metabolism and toxicity. Integr. Biol. (Camb). 3(5), 509–21 (2011) N. Ye, J. Qin, W. Shi et al., Cell-based high content screening using an integrated microfluidic device. Lab Chip 7(12), 1696–1704 (2007)