| Summary: | 碩士 === 國立交通大學 === 生物科技學系 === 107 === ScoreCard, embryoid body formation, teratoma and in vitro differentiation are main methods to identify the pluripotent and differentiation ability of human induced pluripotent stem cells (hiPSC). However, these methods are time consuming and complicated that could not be analyzed easily. In addition, the efficiency of in vitro myocardial directed differentiation is limited by the inability to assess in a short time and individual differences. However, many reference indicate that embryonic development and stem cell fate depend on complex and dynamic cellular microenvironment. The stem cell-extracellular matrix interface is controlled by nanoscale and other physical factors, resulting in nanomorphology that can be used to regulate stem cell behavior. In this study, different size of tantalum oxide nanodot arrays was devised to investigate the effects of various surface nanotopography on hiPSC in the absence of feeder cells or coated extracellular matrices. The qPCR results confirmed that the gene of the pluripotent and three germ layers could be expressed simultaneously on the material after 7 days without chemical induction, and the expression was better than that of the control group. Therefore, the nanodot arrays can effectively amplify its gene expression. In addition, the physical stimulation given by different sizes of nanodot can make different expressions of hiPSC from different sources. According to immunofluorescence staining, this nanodot arrays can promote the differentiation of nerve cells and hepatocytes. However, the results of flow cytometry showed that the differentiation efficiency of traditional myocardial differentiation was different. According to the qPCR results, the pluripotent marker OCT4 on the nanodot arrays was found to be in the flat to 200 nm gene trend if it was associated with the myocardial specific markers NKX2.5 and GATA4. Similarly, it is estimated that the efficiency of traditional myocardial differentiation is better. The cTnT gene expression is not affected by the size of the nanodot, and it can be judged that this material can promote differentiation into late stage of cardiomyocyte. This can be applied not only to hiPSC of different lineages, but also to a screening platform for further development of culture for regenerative medicine.
|
|---|
