Role of Fibroblast Growth Factor 9 in Neurosphere Formation of Adipose-derived Stem Cells

• Main Authors: Shih-YuLu, 陸詩羽
• Other Authors: Chia-Ching Wu
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/ncpa65

碩士 === 國立成功大學 === 細胞生物與解剖學研究所 === 105 === Adipose-derived stem cells (ASCs) were an ideal cell source for stem cell therapy can promote nerve regeneration after injury. The fibroblast growth factor 9 (FGF9) can alter the proliferation and cell-fate decision in stem cells. In my project, we aim to investigate the role of FGF9 during neurosphere formation of ASCs. We used chitosan- coated dish to culture ASCs to induce sphere formation with or without addition of FGF9. The expression of gene was detected by PCR and the protein expression level was observed by Western blot and IF staining. Besides, we investigated the involvement of potential signaling pathway of FGF9 signaling during the sphere formation using specific inhibitors or shRNA. The results showed the spheres formed by ASCs containing both neuronal and glial lineage cells. Addition of FGF9 induced spheres differentiation into Schwann lineage cells. Then, the activation of Akt, ERK signal pathway was decreased after forming the sphere and the inactivation of signal pathway was necessary for sphere formation. However, the phosphorylation of Akt pathway was transiently preserved by FGF9. Furthermore, the involvement of Akt signaling was confirmed by blockage of Akt signaling. The FGFR2 and 3 blocking antibody and shFGFRs was used to block FGFRs to investigate the major FGFR of FGF9 induced Schwann cell differentiation. The expression of Schwann cell markers induced by FGF9 was decreased when knockdown FGFR2. The result suggested that FGF9 could modulate NLC differentiation by activating Akt signaling via binding with FGFR2 during sphere formation. The finding of current study may show the possibility of differentiating ASCs into desired neural lineage by modulating the level of FGF9 during sphere formation.