title: Development of an osteoblast-based 3D continuous-perfusion microfluidic system for drug screening.
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/cm(2) at a flow rate of 0.2 microL/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.
authors: Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, 113-8656, Japan.
authors: Kihoon
authors: K
authors: Jang
authors: Kae
authors: K
authors: Sato
authors: Kazuyo
authors: K
authors: Igawa
authors: Ung-il
authors: UI
authors: Chung
authors: Takehiko
authors: T
authors: Kitamori
mesh_terms: D016475:3T3 Cells; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001331:Automation; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D002454:Cell Differentiation; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics; D010006:Osteoblasts / Q000166:cytology*; D010012:Osteogenesis; D010477:Perfusion; D013314:Stress, Mechanical; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D016212:Transforming Growth Factor beta; D049452:Green Fluorescent Proteins; D000469:Alkaline Phosphatase