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Development of an osteoblast-based 3D continuous-perfusion microfluidic system for drug screening
Kihoon Jang1, Kae Sato, Kazuyo Igawa
1Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, 113-8656, Japan.
Analytical and Bioanalytical Chemistry
|December 18, 2007
Summary
This study presents a microchip system for culturing cells and screening drugs. The system enhances osteoblast differentiation and gene expression, enabling precise monitoring for drug discovery.
Area of Science:
- Biotechnology
- Cell Biology
- Drug Discovery
Background:
- Osteoblast differentiation is crucial for bone health and requires precise monitoring.
- Current methods for assessing osteoblast differentiation are often time-consuming and require large cell numbers.
- Developing efficient cell culture systems for drug screening is essential for identifying bone-promoting compounds.
Purpose of the Study:
- To develop and validate a continuous-perfusion glass microchip system for culturing osteoblastic cells.
- To enable automated, long-term monitoring and supernatant sampling for osteoblast differentiation assays.
- To investigate the effect of shear stress on osteoblast differentiation and gene expression within the microchip system.
Main Methods:
- Utilized mouse Col1a1GFP MC-3T3 E1 osteoblastic cells with a green fluorescent protein (GFP) reporter system.
- Employed a continuous-perfusion glass microchip for automated cell culture and monitoring over 10 days.
- Applied shear stress (0.07 dyne/cm(2)) within 3D microchannels and analyzed alkaline phosphatase (ALP) activity.
Main Results:
- The microchip system successfully cultured cells and monitored differentiation for 10 days.
- Shear stress in the microchannels significantly enhanced GFP expression and osteoblast differentiation.
- Alkaline phosphatase activity was 10-fold higher in the microchip system compared to static 48-well dishes, especially with bone morphogenetic protein 2.
Conclusions:
- The microchip system provides an efficient, precise, and noninvasive method for monitoring osteogenic differentiation.
- This technology facilitates high-throughput drug screening for osteogenic compounds.
- Combining microchip technology with cell-based sensors offers a promising approach for future bone research and drug development.
