[논문]Design, Fabrication, and Application of a Microfluidic Device for Investigating Physical Stress-Induced Behavior in Yeast and Microalgae

Oh, Soojung; Kim, Jangho; Ryu, Hyun Ryul; Lim, Ki-Taek; Chung, Jong Hoon; Jeon, Noo Li

doi:10.5307/jbe.2014.39.3.244

Design, Fabrication, and Application of a Microfluidic Device for Investigating Physical Stress-Induced Behavior in Yeast and Microalgae 원문보기

Journal of biosystems engineering : JBE, v.39 no.3, 2014년, pp.244 - 252

Oh, Soojung (School of Mechanical and Aerospace Engineering, Seoul National University) , Kim, Jangho (Department of Biosystems & Biomaterials Science and Engineering, Seoul National University) , Ryu, Hyun Ryul (School of Mechanical and Aerospace Engineering, Seoul National University) , Lim, Ki-Taek (Department of Biosystems & Biomaterials Science and Engineering, Seoul National University) , Chung, Jong Hoon (Department of Biosystems & Biomaterials Science and Engineering, Seoul National University) , Jeon, Noo Li (School of Mechanical and Aerospace Engineering, Seoul National University)

Abstract ▼ AI-Helper

Purpose: The development of an efficient in vitro cell culture device to process various cells would represent a major milestone in biological science and engineering. However, the current conventional macro-scale in vitro cell culture platforms are limited in their capacity for detailed analysis and determination of cellular behavior in complex environments. This paper describes a microfluidic-based culture device that allows accurate control of parameters of physical cues such as pressure. Methods: A microfluidic device, as a model microbioreactor, was designed and fabricated to culture Saccharomyces cerevisiae and Chlamydomonas reinhardtii under various conditions of physical pressure stimulus. This device was compatible with live-cell imaging and allowed quantitative analysis of physical cue-induced behavior in yeast and microalgae. Results: A simple microfluidic-based in vitro cell culture device containing a cell culture channel and an air channel was developed to investigate physical pressure stress-induced behavior in yeasts and microalgae. The shapes of Saccharomyces cerevisiae and Chlamydomonas reinhardtii could be controlled under compressive stress. The lipid production by Chlamydomonas reinhardtii was significantly enhanced by compressive stress in the microfluidic device when compared to cells cultured without compressive stress. Conclusions: This microfluidic-based in vitro cell culture device can be used as a tool for quantitative analysis of cellular behavior under complex physical and chemical conditions.

주제어

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문제 정의

The fluorescence intensity data still need to be correlated with actual amounts of lipid, but we predict that the enhanced fluorescence signal in stressed cells corresponds to lipid accumulation (Figure 6). The aim of the present study was to provide preliminary results for designing and manipulating microfluidic growth conditions in an in vitro cell culture device. Use of this device has confirmed that we can generate physical stresses that can mimic the specific in vivo environment that occurs in large scale devices, thereby providing us with an efficient strategy for quantitative analysis of living systems in biological science and engineering.

제안 방법

This was also confirmed by the fluorescence microscopy image analysis of GFP-conjugated yeasts cultured under the same conditions (Figure 3(a)). We analyzed the morphologic changes in the yeasts by decovoluting image stacks into 3D images of GFP-conjugated yeasts using IMARIS software. The surfaces of the yeast cells can be monitored in a computerized view with this software (Figure 3(b)).

데이터처리

Data were analyzed using Microsoft Excel 2010, images were processed with ImageJ software, and graphs were drawn with SigmaPlot 10.0. All 3D images were deconvoluted with IMARIS 7.
Student’s t-test was used for statistical analysis using SAS software.

후속연구

The current study focused on the analysis of behavior of yeasts and microalgae in terms of cell shape, but the platform has potential for application to other cell systems, including animal cells, for determining physical stress-induced responses. In conclusion, the results presented in this study collectively demonstrate that this microfluidic-based in vitro cell culture device may be useful as a tool for quantitative analysis of living systems exposed to different physical environments.

참고문헌 (13)

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원문보기 상세보기
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