Epigenetics Involved in the Adhesion and Differentiation of Neural Stem Cell on Fibrous Matrices
| Autor: | shang-Hao Liu, 劉尚豪 |
|---|---|
| Rok vydání: | 2014 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 102 In recent years, neural stem cells (NSCs) therapy holds great importance in tissue engineering. Understanding the control mechanisms of stem cell differentiation for clinical neural therapy is an important research issue. Also, epigenetic regulations of genes were critical in embryonic cell differentiation and fetal development. Therefore, knowledge on the epigenetic mechanisms for NSCs differentiation can provide various and broad applications for regenerative medicine. In addition the effect of a material on the differentiation of NSCs in relation to its epigenetic mechanisms will ensure its safety. The aims of this study are to investigate the effect of the electrospun nanofibrous substrates on the functional genetics and epigenetic profiles of NSC differentiation and identify differentially methylated genes, and to find relevant pathways involved in the NSC differentiation on artificial micro environments. The genomic DNA and RNA are extracted from differentiated and undifferentiated NSCs to confirm the expression of the chosen candidate genes. The genes responsible for the proliferation, growth, and differentiation fate of NSCs are evaluated before and after its differentiation on amino-propyl silica (AP3M) and poly-o-methoxyaniline (POMA). We found eight candidate genes from AP3M and five candidate genes from POMA involved in the NSCs differentiation on three different nanofibrous substrate. In Ingenuity Pathways Analysis (IPA), the gene expressions of eight genes from thirteen candidate genes increased, such as ERBB2, KIF5C, NCAM2, ELMO1, IGF1, NDEL1, NGF and CDH2. To conform the microarray analysis, Bisulfite sequencing PCR (BSP) and real-time PCR (RT-PCR) were verified the DNA methylation of CpG islands and gene expression of these candidate genes. The data demonstrated six genes with methylation changed when NSCs differentiation on nanomaterial. An assessment on the changes of epigenetic mechanism during NSC differentiation in an inorganic nanomaterial will be useful for the medical application of NSCs. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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