Construction and Application of Fluid Shear Stress Bioreactor System

• Main Authors: You-ChingChih, 紀又菁
• Other Authors: Ming-Long Yeh
• Format: Others
• Language: en_US
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/zccdq9

碩士 === 國立成功大學 === 生物醫學工程學系 === 103 === Besides many physiological factors regulating the cells in vivo, mechanical stress also has the obvious influences on cell growth and function. However, it can’t supply these mechanical stimuli through the traditional cell culture process, thus it makes the differences between the cells in vivo and the traditional cell culture. In this study, we constructed a set of auto-controlled bioreactor system in the hood to supply the mechanical stress on cells, and let the adipose-derived stem cells (ADSCs) of the fluid-stimulated experiments under a minimum possibility of contamination through the barrier property of the hood. Meanwhile, it was also convenient for operating experiments in the hood. After stimulating ADSCs under the laminar flow field, calculating the flow field mode and the fluid shear stress (FSS) to ensure the availability of the improved bioreactor system. The examined results showed that the average temperatures were 37.01  0.074, 37.02  0.075, and 37  0.075 ℃ and the average FSS were 3.02  0.001, 3.02  0.003, and 3.02  0.004 dyne/cm2 after during the stimulations of 1, 5, and 10 hours under a steady flow of 3 dyne/cm2. It indicated the temperature and flow rate control exactly achieved high accuracy and stability. However, the calibration of flow rate conducted by the programming language C in microcontroller could conquer the potential operating defects of the peristaltic pump. The programming language C in microcontroller could achieve more precise instantaneous control than the program software control on the Windows Microsoft operating system. Besides, the design of LabVIEW program interface could also supply more selections of the parameters for the newly-designed flow chamber in the future. On the other hand, we clarified the growth condition of ADSCs via the optical microscopy images, the fluorescent images, and the quantitative charts. ADSCs could culture on the polydimethylsiloxane (PDMS) for 24 hours and then separately cultured in two conditions, under the static-state cultivation for 10 hours and the shear stress stimulus of 3 dyne/cm2 for 10 hours. Furthermore, the reaction of ADSCs after stimulation by the laminar flow was observed through the above experiments. The results of optical microscopy images and fluorescent images showed that there were some parts of ADSCs lost after the steady FSS stimuli compared with the static culture group. On the other hands, the elongation with the flow direction of cell cytoskeleton and cell morphology had the insignificantly tendency between the comparisons of the pre-test and post-test in each experimental group and the two groups. Above all of the results in the study, our improved bioreactor system didn’t achieve the expected physiological conditions in cultivating the ADSCs in vitro. We hope that we can modify the bioreactor system to achieve optimal conditions which are closer to the real in vivo condition in the future.