Microprinting of Liver Micro-organ for Drug Metabolism Study

  • Robert C. Chang
  • Kamal Emami
  • Antony Jeevarajan
  • Honglu Wu
  • Wei Sun
Part of the Methods in Molecular Biology book series (MIMB, volume 671)


In their normal in vivo matrix milieu, tissues assume complex well-organized 3D architectures. Therefore, a primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario, in which the precise configuration and composition of cells and bioactive matrix components can establish the well-defined biomimetic microenvironments that promote cell–cell and cell–matrix interactions. With the advent and refinements in microfabricated systems which can present physical and chemical cues to cells in a controllable and reproducible fashion unrealizable with conventional tissue culture, high-fidelity, high-throughput in vitro models are achieved. The convergence of solid freeform fabrication (SFF) technologies, namely microprinting, along with microfabrication techniques, a 3D microprinted micro-organ, can serve as an in vitro platform for cell culture, drug screening, or to elicit further biological insights. This chapter firstly details the principles, methods, and applications that undergird the fabrication process development and adaptation of microfluidic devices for the creation of a drug screening model. This model involves the combinatorial setup of an automated syringe-based, layered direct cell writing microprinting process with soft lithographic micropatterning techniques to fabricate a microscale in vitro device housing a chamber of microprinted 3D micro-organ that biomimics the cell’s natural microenvironment for enhanced performance and functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, 3D cell-encapsulated hydrogel-based tissue constructs are microprinted reproducibly in defined design patterns and biologically characterized for both viability and cell-specific function. Another key facet of the in vivo microenvironment that is recapitulated with the in vitro system is the necessary dynamic perfusion of the 3D microscale liver analog with cells probed for their collective drug metabolic function and suitability as a drug metabolism model.

Key words

Microfluidics Cell printing Tissue engineering Solid freeform fabrication Hydrogels Pharmacokinetics 


  1. 1.
    Chang, R., Nam, J., Holtorf, H., Emami, K., Gonda, S., Jeevarajan, A., Wu, H., Sun, W. (2008) A Case Study on 3D Bioprinted Liver Micro-organ as a Drug Metabolism Model. SME Rapid Technologies/Additive Manufacturing for Medical Applications, Edited by Ola Harrysson, accepted.Google Scholar
  2. 2.
    Powers, M.J., Janigian, D.M., Wack, K.E., Baker, C.S., Stolz, D.B., Griffith, L.G. (2002) Functional behavior of primary rat liver cells in a three-dimensional perfused microarray bioreactor. Tissue Eng 8:499.CrossRefGoogle Scholar
  3. 3.
    Langer, R., Vacanti, J.P. (1993) Tissue engineering. Science 260:920.CrossRefGoogle Scholar
  4. 4.
    Lysaght, M.J., Reyes, J. (2001) The growth of tissue engineering. Tissue Eng 7:485.CrossRefGoogle Scholar
  5. 5.
    Griffith, L.G. (2000) Polymeric biomaterials. Acta Mater 48:263.CrossRefGoogle Scholar
  6. 6.
    Griffith, L.G., Naughton, G. (2002) Tissue engineering – current challenges and expanding opportunities in science. Science 295:1009.CrossRefGoogle Scholar
  7. 7.
    Powers, M.J., Domansky, K., Kaazempur-Mofrad, M.R., Kalezi, A., Capitano, A., Upadhyaya, A., Kurzawski, P., Wack, K.E., Stolz, D.B., Kamm, R., Griffith, L.G. (2002) A microfabricated array bioreactor for perfused 3D liver culture. Biotech Bioeng 78:257.CrossRefGoogle Scholar
  8. 8.
    Ghanem, A., Shuler, M.L. (2000) Combining cell culture analogue reactor designs and PBPK models to probe mechanisms of naphthalene toxicity. Biotechnol Prog 16:334.CrossRefGoogle Scholar
  9. 9.
    Shuler, M.L., Ghanem, A., Quick, D., Wong, M.C., Miller, P. (1996) A self-regulating cell culture analog device to mimic animal and human toxicological responses. Biotechnol Bioeng 52:45.CrossRefGoogle Scholar
  10. 10.
    Viravaidya, K., Sin A., Shuler M.L. (2004) Development of a microscale cell culture analog to probe naphthalene toxicity. Biotechnol Prog 20:316.CrossRefGoogle Scholar
  11. 11.
    Bratten, C.D.T., Cobbold, P.H., Cooper, J.M. (1998) Single-cell measurements of purine release using a micromachined electroanalytical sensor. Anal Chem 70:1164.CrossRefGoogle Scholar
  12. 12.
    Chen, P., Xu, B., Tokranova, N., Feng, X., Castracane, J., Gillis, K.D. (2003) Amperometric detection of quantal catecholamine secretion from individual cells on micromachined silicon chips. Anal Chem 75:518.CrossRefGoogle Scholar
  13. 13.
    Meyvantsson, I., Beebe, D.J. (2008) Cell culture models in microfluidic systems. Ann Rev Anal Chem 1:423.CrossRefGoogle Scholar
  14. 14.
    Borenstein, J.T., Weinberg, E.J., Orrick, B.K., Sundback, C., Kaazempur-Mofrad, M.R., Vacanti, J.P. (2007) Microfabrication of three-dimensional engineered scaffolds. Tissue Eng 13(8):1837.CrossRefGoogle Scholar
  15. 15.
    Holmes, T.C., de Lacalle, S., Su, X., Liu, G., Rich, A., and Zhang, S. (2000) Extensive neurite outgrowth and active synapse formation on self-assembling peptide scaffolds. Proc Nat Acad Sci USA 97:6728.CrossRefGoogle Scholar
  16. 16.
    Semino, C.E., Merok, J.R., Crane, G.G., Panagiotakos, G., Zhang, S. (2003) Functional differentiation of hepatocytelike spheroid structures from putative liver progenitor cells in three-dimensional peptide scaffolds. Differentiation 71:262.CrossRefGoogle Scholar
  17. 17.
    Weaver, V.M., Petersen, O.W., Wang, F., Larabell, C.A., Briand, P., Damsky, C. (1997) Reversion of the malignant phenotype of human breast cells in three-dimensional culture and in vivo by integrin blocking antibodies. J Cell Biol 137(1):231.CrossRefGoogle Scholar
  18. 18.
    Chang, R., Nam, J., Sun, W. (2008) Effects of dispensing pressure and nozzle diameter on cell survival from solid freeform fabrication-based direct cell writing. Tissue Eng 14(1):41.CrossRefGoogle Scholar
  19. 19.
    Chang, R., Nam, J., Sun, W. (2008) Direct cell writing of 3D micro-organ for in vitro pharmacokinetic model. Tissue Eng C 14(2):157.CrossRefGoogle Scholar
  20. 20.
    Chang, R., Nam, J., Holtorf, H., Emami, K., Gonda, S., Jeevarajan, A., Wu, H., Sun, W. (2008) Bioprinting of micro-organ tissue analog for drug metabolism study. 37th COSPAR Scientific Assembly, Montreal, Canada, July 13–20, 2008.Google Scholar
  21. 21.
    Khalil, S., Sun, W. (2007) Biopolymer deposition for freeform fabrication of hydrogel tissue constructs. Mater Sci Eng C 27:478.CrossRefGoogle Scholar
  22. 22.
    Khalil, S., Nam, J., Sun, W. (2005) Multi-nozzle deposition for construction of 3D biopolymer tissue scaffolds. Rapid Prototyping J 11:9.CrossRefGoogle Scholar
  23. 23.
    Knight, B., Laukaitis, C., Akhtar, N., Hotchin, N.A., Edlund, M., Horwitz, A.F. (2000) Visualizing muscle cell migration. Curr Biol 10:576.CrossRefGoogle Scholar
  24. 24.
    Mooney, D.J., Sano, K., Kaufmann, P.M., Majahod, K., Schloo, B., Vacanti, J.P., Langer, R. (1996) Long-term culture of hepatocytes transplanted on biodegradable polymer sponges. J Biomed Mater Res 37:413.CrossRefGoogle Scholar
  25. 25.
    Patz, T.M., Doraiswamy, A., Narayan, R.J. (2006) Three-dimensional direct writing of B35 neuronal cells. J Biomed Mater Res B 78(1):124.Google Scholar
  26. 26.
    Roskelley, C.D., Desprez, P.Y., Bissell, M.J. (1994) Extracellular matrix-dependent tissue specific gene expression in mammary epithelial cells requires both physical and biochemical signal transduction. Proc Nat Acad Sci USA 91:12378.CrossRefGoogle Scholar
  27. 27.
    Abbot, A. (2003) Biology’s new dimension. Nature 424:870.CrossRefGoogle Scholar
  28. 28.
    Sahai, E., Marshall, C.J. (2003) Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signalling and extracellular proteolysis. Nat Cell Biol 5(8):711.CrossRefGoogle Scholar
  29. 29.
    Wolf, K., Mazo, I., Leung, H., Engelke, K., von Andrian, U.H., Deryugina, E.I. (2003) Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis. J Cell Biol 160(2):267.CrossRefGoogle Scholar
  30. 30.
    El-Ali, J., Sorger, P.K. and Jensen, K.F. (2006) Cells on chips. Nature 442:403.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Robert C. Chang
    • 1
  • Kamal Emami
    • 2
  • Antony Jeevarajan
    • 2
  • Honglu Wu
    • 2
  • Wei Sun
    • 1
  1. 1.Department of Mechanical Engineering and MechanicsDrexel UniversityPhiladelphiaUSA
  2. 2.Radiation Physics LaboratoryNASA Johnson Space CenterHoustonUSA

Personalised recommendations