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.
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