Analytical and Bioanalytical Chemistry

, Volume 390, Issue 3, pp 825–832

Development of an osteoblast-based 3D continuous-perfusion microfluidic system for drug screening

Authors

  • Kihoon Jang
    • Department of Applied Chemistry, Graduate School of EngineeringThe University of Tokyo
  • Kae Sato
    • Department of Applied Chemistry, Graduate School of EngineeringThe University of Tokyo
    • Center for NanoBio IntegrationThe University of Tokyo
  • Kazuyo Igawa
    • Division of Tissue EngineeringThe University of Tokyo Hospital
  • Ung-il Chung
    • Center for NanoBio IntegrationThe University of Tokyo
    • Division of Tissue EngineeringThe University of Tokyo Hospital
    • Department of Bioengineering, School of EngineeringThe University of Tokyo
    • Department of Applied Chemistry, Graduate School of EngineeringThe University of Tokyo
    • Core Research for Evolutional Science and TechnologyJapan Science and Technology Agency
    • Center for NanoBio IntegrationThe University of Tokyo
Original Paper

DOI: 10.1007/s00216-007-1752-7

Cite this article as:
Jang, K., Sato, K., Igawa, K. et al. Anal Bioanal Chem (2008) 390: 825. doi:10.1007/s00216-007-1752-7

Abstract

In this work, we demonstrated that biological cells could be cultured in a continuous-perfusion glass microchip system for drug screening. We used mouse Col1a1GFP MC-3T3 E1 osteoblastic cells, which have a marker gene system expressing green fluorescent protein (GFP) under the control of osteoblast-specific promoters. With our microchip-based cell culture system, we realized automated long-term monitoring of cells and sampling of the culture supernatant system for osteoblast differentiation assay using a small number of cells. The system successfully monitored cells for 10 days. Under the 3D microchannel condition, shear stress (0.07 dyne/cm2 at a flow rate of 0.2 μL/min) was applied to the cells and it enhanced the GFP expression and differentiation of the osteoblasts. Analysis of alkaline phosphatase (ALP), which is an enzyme marker of osteoblasts, supported the results of GFP expression. In the case of differentiation medium containing bone morphogenetic protein 2, we found that ALP activity in the culture supernatant was enhanced 10 times in the microchannel compared with the static condition in 48-well dishes. A combined system of a microchip and a cell-based sensor might allow us to monitor osteogenic differentiation easily, precisely, and noninvasively. Our system can be applied in high-throughput drug screening assay for discovering osteogenic compounds.

Keywords

BioassaysCell systemsMicrofluidicsOsteoblastShear stressDifferentiation

Copyright information

© Springer-Verlag 2007