Abstract
Microtechnology provides a new approach for reproducing the in vivo environment in vitro. Mimicking the microenvironment of the natural tissues allows cultured cells to behave in a more authentic manner, and gives researchers more realistic platforms to study biological systems. In this review article, we discuss the physiochemical aspects of in vivo cellular microenvironment, and relevant technologies that can be used to mimic those aspects. Secondly we identify the core methods used in microtechnology for biomedical applications. Finally we examine the recent application areas of microtechnology, with a focus on reproducing the functions of specific organs, or whole-body response such as homeostasis or metabolism-dependent toxicity of drugs. These new technologies enable researchers to ask and answer questions in a manner that has not been possible with conventional, macroscale technologies.
Similar content being viewed by others
References
Allen, J. W., S. R. Khetani, and S. N. Bhatia. In vitro zonation and toxicity in a hepatocyte bioreactor. Toxicol. Sci. 84(1):110–119, 2005.
Artursson, P., K. Palm, and K. Luthman. Caco-2 monolayers in experimental and theoretical predictions of drug transport. Adv. Drug Deliv. Rev. 46(1–3):27–43, 2001.
Baudoin, R., A. Corlu, L. Griscom, C. Legallais, and E. Leclerc. Trends in the development of microfluidic cell biochips for in vitro hepatotoxicity. Toxicol. In Vitro 21(4):535–544, 2007.
Bennett, M. R., and J. Hasty. Microfluidic devices for measuring gene network dynamics in single cells. Nat. Rev. Genet. 10(9):628–638, 2009.
Bergman, R. N. Orchestration of glucose homeostasis: from a small acorn to the California oak. Diabetes 56(6):1489–1501, 2007.
Bhatia, S. N., M. L. Yarmush, and M. Toner. Controlling cell interactions by micropatterning in co-cultures: hepatocytes and 3T3 fibroblasts. J. Biomed. Mater. Res. 34(2):189–199, 1997.
Brandon, E. F., C. D. Raap, I. Meijerman, J. H. Beijnen, and J. H. Schellens. An update on in vitro test methods in human hepatic drug biotransformation research: pros and cons. Toxicol. Appl. Pharmacol. 189(3):233–246, 2003.
Burgess, K. A., H. H. Hu, W. R. Wagner, and W. J. Federspiel. Towards microfabricated biohybrid artificial lung modules for chronic respiratory support. Biomed. Microdevices 11(1):117–127, 2009.
Camp, J. P., T. Stokol, and M. L. Shuler. Fabrication of a multiple-diameter branched network of microvascular channels with semi-circular cross-sections using xenon difluoride etching. Biomed. Microdevices 10(2):179–186, 2008.
Cukierman, E., R. Pankov, D. R. Stevens, and K. M. Yamada. Taking cell–matrix adhesions to the third dimension. Science 294(5547):1708–1712, 2001.
De Smet, K., T. Bruning, M. Blaszkewicz, H. M. Bolt, A. Vercruysse, and V. Rogiers. Biotransformation of trichloroethylene in collagen gel sandwich cultures of rat hepatocytes. Arch. Toxicol. 74(10):587–592, 2000.
Dittrich, P. S., and A. Manz. Lab-on-a-chip: microfluidics in drug discovery. Nat. Rev. Drug Discov. 5(3):210–218, 2006.
Douville, N. J., P. Zamankhan, Y. C. Tung, R. Li, B. L. Vaughan, C. F. Tai, J. White, P. J. Christensen, J. B. Grotberg, and S. Takayama. Combination of fluid and solid mechanical stresses contribute to cell death and detachment in a microfluidic alveolar model. Lab Chip 11(4):609–619, 2011.
Fidkowski, C., M. R. Kaazempur-Mofrad, J. Borenstein, J. P. Vacanti, R. Langer, and Y. Wang. Endothelialized microvasculature based on a biodegradable elastomer. Tissue Eng. 11(1–2):302–309, 2005.
Geckil, H., F. Xu, X. Zhang, S. Moon, and U. Demirci. Engineering hydrogels as extracellular matrix mimics. Nanomedicine (Lond.) 5(3):469–484, 2010.
Golden, A. P., and J. Tien. Fabrication of microfluidic hydrogels using molded gelatin as a sacrificial element. Lab Chip 7(6):720–725, 2007.
Guillaume-Gentil, O., M. Gabi, M. Zenobi-Wong, and J. Voros. Electrochemically switchable platform for the micro-patterning and release of heterotypic cell sheets. Biomed. Microdevices 13(1):221–230, 2011.
Guillotin, B., and F. Guillemot. Cell patterning technologies for organotypic tissue fabrication. Trends Biotechnol. 29(4):183–190, 2011.
Guzzardi, M. A., C. Domenici, and A. Ahluwalia. Metabolic control through hepatocyte and adipose tissue cross-talk in a multicompartmental modular bioreactor. Tissue Eng. A 17(11–12):1635–1642, 2011.
Honarmandi, P., H. Lee, M. J. Lang, and R. D. Kamm. A microfluidic system with optical laser tweezers to study mechanotransduction and focal adhesion recruitment. Lab Chip 11(4):684–694, 2011.
Hosmane, S., A. Fournier, R. Wright, L. Rajbhandari, R. Siddique, I. H. Yang, K. T. Ramesh, A. Venkatesan, and N. Thakor. Valve-based microfluidic compression platform: single axon injury and regrowth. Lab Chip 11(22):3888–3895, 2011.
Huh, D., B. D. Matthews, A. Mammoto, M. Montoya-Zavala, H. Y. Hsin, and D. E. Ingber. Reconstituting organ-level lung functions on a chip. Science 328(5986):1662–1668, 2010.
Humes, H. D., W. H. Fissell, and K. Tiranathanagul. The future of hemodialysis membranes. Kidney Int. 69(7):1115–1119, 2006.
Ismagilov, R. F., and M. M. Maharbiz. Can we build synthetic, multicellular systems by controlling developmental signaling in space and time? Curr. Opin. Chem. Biol. 11(6):604–611, 2007.
Janmey, P. A., and C. A. McCulloch. Cell mechanics: integrating cell responses to mechanical stimuli. Annu. Rev. Biomed. Eng. 9:1–34, 2007.
Jeong, G. S., S. Chung, C. B. Kim, and S. H. Lee. Applications of micromixing technology. Analyst 135(3):460–473, 2010.
Kang, J. H., Y. C. Kim, and J. K. Park. Analysis of pressure-driven air bubble elimination in a microfluidic device. Lab Chip 8(1):176–178, 2008.
Keenan, T. M., and A. Folch. Biomolecular gradients in cell culture systems. Lab Chip 8(1):34–57, 2008.
Khademhosseini, A., and R. Langer. Microengineered hydrogels for tissue engineering. Biomaterials 28(34):5087–5092, 2007.
Khaleque, T., S. Abu-Salih, J. R. Saunders, and W. Moussa. Experimental methods of actuation, characterization and prototyping of hydrogels for bioMEMS/NEMS applications. J. Nanosci. Nanotechnol. 11(3):2470–2479, 2011.
Khetani, S. R., and S. N. Bhatia. Microscale culture of human liver cells for drug development. Nat. Biotechnol. 26(1):120–126, 2008.
Kim, S., H. J. Kim, and N. L. Jeon. Biological applications of microfluidic gradient devices. Integr. Biol. (Camb.) 2(11-12):584–603, 2010.
Kim, L., M. D. Vahey, H. Y. Lee, and J. Voldman. Microfluidic arrays for logarithmically perfused embryonic stem cell culture. Lab Chip 6(3):394–406, 2006.
Kim, S. J., F. Wang, M. A. Burns, and K. Kurabayashi. Temperature-programmed natural convection for micromixing and biochemical reaction in a single microfluidic chamber. Anal. Chem. 81(11):4510–4516, 2009.
Lee, K., C. Kim, B. Ahn, R. Panchapakesan, A. R. Full, L. Nordee, J. Y. Kang, and K. W. Oh. Generalized serial dilution module for monotonic and arbitrary microfluidic gradient generators. Lab Chip 9(5):709–717, 2009.
Lee, M. Y., R. A. Kumar, S. M. Sukumaran, M. G. Hogg, D. S. Clark, and J. S. Dordick. Three-dimensional cellular microarray for high-throughput toxicology assays. Proc. Natl. Acad. Sci. USA 105(1):59–63, 2008.
Lee, S. H., D. van Noort, J. Y. Lee, B. T. Zhang, and T. H. Park. Effective mixing in a microfluidic chip using magnetic particles. Lab Chip. 9(3):479–482, 2009.
Lehmann, A. D., N. Daum, M. Bur, C. M. Lehr, P. Gehr, and B. M. Rothen-Rutishauser. An in vitro triple cell co-culture model with primary cells mimicking the human alveolar epithelial barrier. Eur. J. Pharm. Biopharm. 77(3):398–406, 2011.
Leonard, E. F., S. Cortell, and N. G. Vitale. Membraneless dialysis—is it possible? Contrib. Nephrol. 149:343–353, 2005.
Li Jeon, N., H. Baskaran, S. K. Dertinger, G. M. Whitesides, L. Van de Water, and M. Toner. Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device. Nat. Biotechnol. 20(8):826–830, 2002.
Lu, H., L. Y. Koo, W. M. Wang, D. A. Lauffenburger, L. G. Griffith, and K. F. Jensen. Microfluidic shear devices for quantitative analysis of cell adhesion. Anal. Chem. 76(18):5257–5264, 2004.
Lucchetta, E. M., J. H. Lee, L. A. Fu, N. H. Patel, and R. F. Ismagilov. Dynamics of Drosophila embryonic patterning network perturbed in space and time using microfluidics. Nature 434(7037):1134–1138, 2005.
Ma, B., G. Zhang, J. Qin, and B. Lin. Characterization of drug metabolites and cytotoxicity assay simultaneously using an integrated microfluidic device. Lab Chip 9(2):232–238, 2009.
Mahler, G. J., M. B. Esch, R. P. Glahn, and M. L. Shuler. Characterization of a gastrointestinal tract microscale cell culture analog used to predict drug toxicity. Biotechnol. Bioeng. 104(1):193–205, 2009.
Mahler, G. J., M. L. Shuler, and R. P. Glahn. Characterization of Caco-2 and HT29-MTX cocultures in an in vitro digestion/cell culture model used to predict iron bioavailability. J. Nutr. Biochem. 20(7):494–502, 2009.
McGuigan, A. P., and M. V. Sefton. Vascularized organoid engineered by modular assembly enables blood perfusion. Proc. Natl. Acad. Sci. USA 103(31):11461–11466, 2006.
Meyvantsson, I., J. W. Warrick, S. Hayes, A. Skoien, and D. J. Beebe. Automated cell culture in high density tubeless microfluidic device arrays. Lab Chip 8(5):717–724, 2008.
Milosevic, N., H. Schawalder, and P. Maier. Kupffer cell-mediated differential down-regulation of cytochrome P450 metabolism in rat hepatocytes. Eur. J. Pharmacol. 368(1):75–87, 1999.
Moon, J. J., and J. L. West. Vascularization of engineered tissues: approaches to promote angio-genesis in biomaterials. Curr. Top Med. Chem. 8(4):300–310, 2008.
Morier, P., C. Vollet, P. E. Michel, F. Reymond, and J. S. Rossier. Gravity-induced convective flow in microfluidic systems: electrochemical characterization and application to enzyme-linked immunosorbent assay tests. Electrophoresis 25(21–22):3761–3768, 2004.
Mrksich, M., L. E. Dike, J. Tien, D. E. Ingber, and G. M. Whitesides. Using microcontact printing to pattern the attachment of mammalian cells to self-assembled monolayers of alkanethiolates on transparent films of gold and silver. Exp. Cell Res. 235(2):305–313, 1997.
Musi, N., and L. J. Goodyear. Insulin resistance and improvements in signal transduction. Endocrine 29(1):73–80, 2006.
Nahmias, Y., F. Berthiaume, and M. L. Yarmush. Integration of technologies for hepatic tissue engineering. Adv. Biochem. Eng. Biotechnol. 103:309–329, 2007.
Orr, D. E., and K. J. Burg. Design of a modular bioreactor to incorporate both perfusion flow and hydrostatic compression for tissue engineering applications. Ann. Biomed. Eng. 36(7):1228–1241, 2008.
Park, T. H., and M. L. Shuler. Integration of cell culture and microfabrication technology. Biotechnol. Prog. 19(2):243–253, 2003.
Pelkonen, O., and M. Turpeinen. In vitro–in vivo extrapolation of hepatic clearance: biological tools, scaling factors, model assumptions and correct concentrations. Xenobiotica 37(10–11):1066–1089, 2007.
Ramello, C., P. Paullier, A. Ould-Dris, M. Monge, C. Legallais, and E. Leclerc. Investigation into modification of mass transfer kinetics by acrolein in a renal biochip. Toxicol. In Vitro 25(5):1123–1131, 2011.
Saadi, W., S. W. Rhee, F. Lin, B. Vahidi, B. G. Chung, and N. L. Jeon. Generation of stable concentration gradients in 2D and 3D environments using a microfluidic ladder chamber. Biomed. Microdevices 9(5):627–635, 2007.
Schiele, N. R., D. T. Corr, Y. Huang, N. A. Raof, Y. Xie, and D. B. Chrisey. Laser-based direct-write techniques for cell printing. Biofabrication 2(3):032001, 2010.
Shah, R. K., and A. L. London, Laminar Flow Forced Convection in Ducts: A Source Book for Compact Heat Exchanger Analytical Data. Advances in Heat Transfer Supplement. New York: Academic Press, xiv, 477 pp., 1978.
Sharma, R. I., and J. G. Snedeker. Biochemical and biomechanical gradients for directed bone marrow stromal cell differentiation toward tendon and bone. Biomaterials 31(30):7695–7704, 2010.
Sin, A., K. C. Chin, M. F. Jamil, Y. Kostov, G. Rao, and M. L. Shuler. The design and fabrication of three-chamber microscale cell culture analog devices with integrated dissolved oxygen sensors. Biotechnol. Prog. 20(1):338–345, 2004.
Sivaraman, A., J. K. Leach, S. Townsend, T. Iida, B. J. Hogan, D. B. Stolz, R. Fry, L. D. Samson, S. R. Tannenbaum, and L. G. Griffith. A microscale in vitro physiological model of the liver: predictive screens for drug metabolism and enzyme induction. Curr. Drug Metab. 6(6):569–591, 2005.
Skelley, A. M., and J. Voldman. An active bubble trap and debubbler for microfluidic systems. Lab Chip 8(10):1733–1737, 2008.
Stoltz, J. F., S. Muller, A. Kadi, V. Decot, P. Menu, and D. Bensoussan. Introduction to endothelial cell biology. Clin. Hemorheol. Microcirc. 37(1–2):5–8, 2007.
Stroock, A. D., and C. Fischbach. Microfluidic culture models of tumor angiogenesis. Tissue Eng. A 16(7):2143–2146, 2010.
Sundararaghavan, H. G., G. A. Monteiro, B. L. Firestein, and D. I. Shreiber. Neurite growth in 3D collagen gels with gradients of mechanical properties. Biotechnol. Bioeng. 102(2):632–643, 2009.
Sung, J. H., M. B. Esch, and M. L. Shuler. Integration of in silico and in vitro platforms for pharmacokinetic-pharmacodynamic modeling. Expert Opin. Drug Metab. Toxicol. 6(9):1063–1081, 2010.
Sung, J. H., C. Kam, and M. L. Shuler. A microfluidic device for a pharmacokinetic-pharmacodynamic (PK-PD) model on a chip. Lab Chip 10(4):446–455, 2010.
Sung, J. H., and M. L. Shuler. A micro cell culture analog (microCCA) with 3-D hydrogel culture of multiple cell lines to assess metabolism-dependent cytotoxicity of anti-cancer drugs. Lab Chip 9(10):1385–1394, 2009.
Sung, J. H., and M. L. Shuler. Prevention of air bubble formation in a microfluidic perfusion cell culture system using a microscale bubble trap. Biomed. Microdevices 11(4):731–738, 2009.
Sung, J. H., J. Yu, D. Luo, M. L. Shuler, and J. C. March. Microscale 3-D hydrogel scaffold for biomimetic gastrointestinal (GI) tract model. Lab Chip 11(3):389–392, 2011.
Toepke, M. W., and D. J. Beebe. PDMS absorption of small molecules and consequences in microfluidic applications. Lab Chip 6(12):1484–1486, 2006.
Torisawa, Y. S., B. Mosadegh, G. D. Luker, M. Morell, K. S. O’Shea, and S. Takayama. Microfluidic hydrodynamic cellular patterning for systematic formation of co-culture spheroids. Integr. Biol. (Camb.) 1(11–12):649–654, 2009.
Tsang, V. L., and S. N. Bhatia. Fabrication of three-dimensional tissues. Adv. Biochem. Eng. Biotechnol. 103:189–205, 2007.
Tschumperlin, D. J., and S. S. Margulies. Equibiaxial deformation-induced injury of alveolar epithelial cells in vitro. Am. J. Physiol. 275(6 Pt 1):L1173–L1183, 1998.
Tse, J. R., and A. J. Engler. Stiffness gradients mimicking in vivo tissue variation regulate mesenchymal stem cell fate. PLoS One 6(1):e15978, 2011.
van Midwoud, P. M., M. T. Merema, E. Verpoorte, and G. M. Groothuis. A microfluidic approach for in vitro assessment of interorgan interactions in drug metabolism using intestinal and liver slices. Lab Chip 10(20):2778–2786, 2010.
Vickerman, V., J. Blundo, S. Chung, and R. Kamm. Design, fabrication and implementation of a novel multi-parameter control microfluidic platform for three-dimensional cell culture and real-time imaging. Lab Chip 8(9):1468–1477, 2008.
Viravaidya, K., A. Sin, and M. L. Shuler. Development of a microscale cell culture analog to probe naphthalene toxicity. Biotechnol. Prog. 20(1):316–323, 2004.
Vozzi, F., J. M. Heinrich, A. Bader, and A. D. Ahluwalia. Connected culture of murine hepatocytes and HUVEC in a multicompartmental bioreactor. Tissue Eng. A 15(6):1291–1299, 2009.
Wang, F., V. M. Weaver, O. W. Petersen, C. A. Larabell, S. Dedhar, P. Briand, R. Lupu, and M. J. Bissell. Reciprocal interactions between beta1-integrin and epidermal growth factor receptor in three-dimensional basement membrane breast cultures: a different perspective in epithelial biology. Proc. Natl. Acad. Sci. USA 95(25):14821–14826, 1998.
Whitesides, G. M. The origins and the future of microfluidics. Nature 442(7101):368–373, 2006.
Wnek, G. E., and G. L. Bowlin. Encyclopedia of Biomaterials and Biomedical Engineering. New York: Marcel Dekker, 2004.
Wright, D., B. Rajalingam, S. Selvarasah, M. R. Dokmeci, and A. Khademhosseini. Generation of static and dynamic patterned co-cultures using microfabricated parylene-C stencils. Lab Chip 7(10):1272–1279, 2007.
Xiao, Y., and G. A. Truskey. Effect of receptor-ligand affinity on the strength of endothelial cell adhesion. Biophys. J. 71(5):2869–2884, 1996.
Young, E. W., and D. J. Beebe. Fundamentals of microfluidic cell culture in controlled microenvironments. Chem. Soc. Rev. 39(3):1036–1048, 2010.
Young, E. W., and C. A. Simmons. Macro- and microscale fluid flow systems for endothelial cell biology. Lab Chip 10(2):143–160, 2010.
Zhang, W., S. Lin, C. Wang, J. Hu, C. Li, Z. Zhuang, Y. Zhou, R. A. Mathies, and C. J. Yang. PMMA/PDMS valves and pumps for disposable microfluidics. Lab Chip 9(21):3088–3094, 2009.
Zheng, Y., W. Dai, and H. Wu. A screw-actuated pneumatic valve for portable, disposable microfluidics. Lab Chip 9(3):469–472, 2009.
Acknowledgments
This work was supported by Army Corp of Engineers (CERL, W9132T-07), Nanobiotechnology center (NBTC), National Research Foundation of Korea (NRF, Grant no. 2011-0013862), Hongik University new faculty research support fund, and 2011 Hongik University Research Fund.
Author information
Authors and Affiliations
Corresponding author
Additional information
Associate Editor Tingrui Pan oversaw the review of this article.
Rights and permissions
About this article
Cite this article
Sung, J.H., Shuler, M.L. Microtechnology for Mimicking In Vivo Tissue Environment. Ann Biomed Eng 40, 1289–1300 (2012). https://doi.org/10.1007/s10439-011-0491-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10439-011-0491-2