Abstract
Nowadays, most people are focusing on their works and ignoring the importance of diet habit and sports, which causes a lot of diseases. The storage capability of kidney is very important to human bodies. However symptoms would appear only when the kidneys get damaged seriously. Therefore, any early symptoms cannot be ignored. It is important to do a careful examination and follow the diagnosis until the body gets totally cured; otherwise it may cause kidney inflammation. In the statistics of 2007, kidney disease was ranked up to number eight in Taiwan. According to the statistical data published by USRDS in 2013, the incidence of end-stage kidney disease has ranked from first place to third place. Although the patients of end-stage kidney disease decreased, the population of hemodialysis still rises up because it can extend the life effectively. While hemodialysis machines are used to help patients remove the impurities in the blood, it also means that the function of the kidney cannot be repaired, and patients have to rely on it for the rest of their lives. Therefore, an early detection and regular treatments of the kidney disease may avoid the situation of hemodialysis. The purpose of this master study is to establish a triplicate microfluidic immunoassay system mimicking the kidney immune response in vitro microenvironment for the studies of kidney disease. Therefore, we use MEMS technology to produce a kidney immunoassay chip, combine narrow-gap channel to define the position of the cells, and increase the flow resistance to generate the concentration gradient diffusion. In this research, we use 1 ng/ml, 5 ng/ml and 10 ng/ml three different concentration of Escherichia coli lipopolysaccharide (E.coli LPS) to stimulate renal tubular epithelial cells (HK-2). Then, we use ELISA to detect the cytokine which secret by HK-2 and decide to use 10 ng/ml E.coli LPS in following experiment. After HK-2 cells are stimulated by 10 ng/ml E.coli LPS, it will secret cytokine to affect the human acute monocyte leukemia cell line (THP-1) and we find out that the THP-1 will be differentiated into macrophage and have some defensive action to protect the HK-2. In this study, our design can ensure the accuracy of the experiments and increase the flow resistance to generate concentration gradient diffusion. This microfluidic chip also has the features of low-dose specimen requirement, low cost and simultaneous bio-experiments operated in an array form.