Development of a Multi-Compartment Micro-Cell Culture Device as a Future On-Chip Human: Fabrication of a Three-Compartment Device and Immobilization of Mature Rat Adipocytes for the Evaluation of Chemical Distributions
Hidenari Nakayama*, Hiroshi Kimura*, Kikuo Komori*,
Teruo Fujii*, and Yasuyuki Sakai*,**
*Institute of Industrial Science (IIS), University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan
**Center for Disease Biology and Integrative Medicine (CBIM), University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Absorption, distribution, metabolism and excretion (ADME) are important in estimating the influence of chemicals on health. Distribution among internal organs is difficult to estimate without animal experiments. In vitro development targets the perfusion culturing of multiple cells derived from organs such as the liver, stomach, small intestines, and kidney. Using cheep, easily molded polydimethylsiloxane (PDMS), we produced a cell-culture microdevice having three compartments to determine the kinetic distribution of hydrophobic chemicals imaged by fluorescent imaging based on the presence of mature rat adipocytes. The disposable device uses liquid feeding using a magnetic stirrer. Separate and complete perfusion modes are easily changed by a valve after nurturing organ-derived cells in the device under separate conditions, enabling kinetic evaluation. To stabilize, disperse, and fix mature adipocytes whose specific gravity is lower than the culture solution, nonwoven fabric is used as a three-dimensional scaffold. When fluoranthene, a fluorescent hydrophobic chemical, is added during perfusion culturing, fluoranthene selectively accumulates in a fat compartment after six hours in a device to which adipocytes are added, enabling in vitro determination of hydrophobic chemical accumulation determining the distribution of chemicals in adipocytes. By introducing cells of target organs and metabolic organs in to other compartments, the device is extremely effective in experimentally determining the ADME of chemicals and the development of toxicity in vitro.
-  B. Blaauboer, “The integration on physico-chemical properties, in vitro-derived toxicity data and physiologically based kinetic and dynamic as modeling a tool in hazard and risk assessment,” A commentary, Toxicology Letters, Vol.138, No.2, pp. 161-171, 2003.
-  K. Viravaidya, A. Sin, and M. L. Shuler, “Development of a microscale cell culture analog to probe naphthalene toxicity,” Biotechnology Progress, 20, pp. 316-323, 2004.
-  K. Viravaidya and M. L. Shuler, “Prediction of naphthalene bioaccumulation using an adipocyte cell line model,” Biotechnology Progress, 18, pp. 174-181, 2002.
-  K. Viravaidya and M. L. Shuler, “Incorporation of 3T3-L1 cells to mimic bioaccumulation in a microscale cell culture analog device for toxicity studies,” Biotechnology Progress, 20, pp. 590-597, 2004.
-  HmREL Corporation home page,
-  E. Leclerc, Y. Sakai, and T. Fujii, “Cell culture in 3-dimensional microfluidic structure of PDMS (polydimethylsiloxane),” Biomedical Microdevices, 5(2), pp. 109-114, 2003.
-  E. Leclerc, Y. Sakai, and T. Fujii, “Microfluidic PDMS (Polydimethylsiloxane) Bioreactor for Large-Scale Culture of Hepatocytes,” Biotechnology Progress, 20(3), pp. 750-755, 2004.
-  L. H. Lu, K. S. Ryu, and C. Liu, “A magnetic microstirrer and array for microfluidic mixing. Journal of microelectromechanical systems, 11(5), pp. 462-469, 2002.
-  P. K. Yuen, G. Li, Y. Bao, and U. R. Muller, “Microfluidic devices for fluidic circulation and mixing improve hybridization signal intensity on DNA arrays,” Lab on a chip, 3, pp. 46-50, 2003.
-  K. S. Ryu, K. Shaikh, and C. Liu, “Micro magnetic stir-bars integrated in parylene surface-micromachined channels for mixing and pumping,” 7th Int. Conf. on miniaturized chemical and biochemical analysis system, pp. 635-638, 2003.
-  K. S. Ryu, K. Shaikh, E. Goluch, Z. Fan, and C. Liu, “Micro magnetic stir-bar mixer integrated with parylene microfluidic channels,” Lab on a chip, 4, pp. 608-613, 2004.
-  A. K. Agarwal, S. S. Sridharamurthy, D. J. Beebe, and H. Jiang, “Programmable autonomous micromixers and micropumps,” Journal of microelectromechanical systems, 14(6), pp. 1409-1421, 2005.
-  K. Tsuchiya, T. Yoshimoto, Y. Hirono, T. Tateno, T. Sugiyama, and Y. Hirata, “Angiotensin II induces monocyte chemoattractant protein-1 expression via nuclear factor-B-dependent pathway in rat preadipocyte. American journal of physiology,” Endocrinology and metabolism, p. 291, E771-E778, 2006.
-  H. Sugihara, N. Yonemitsu, S. Miyabara, and S. Toda, “Proliferation of unilocular fat cells in the primary culture,” Journal of Lipid Research, 28, pp. 1038-1045, 1987.
This article is published under a Creative Commons Attribution-NoDerivatives 4.0 Internationa License.
Copyright© 2007 by Fuji Technology Press Ltd. and Japan Society of Mechanical Engineers. All right reserved.