Analysis of Neuronal Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells

• Main Authors: Wan-Chen Wang, 王琬禎
• Other Authors: Wen-Chien Lee
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/61848741916559365570

碩士 === 國立中正大學 === 化學工程所 === 97 === The unique properties of stemness and multipotent differentiation capacity of plastic-adherent human bone marrow mesenchymal stem cells (hMSCs) have been studied extensively. Moreover, recent research indicates that the hMSCs not only can differentiate into mesodermal cell lineages but also can transdifferentiate into cell types of the other germ layers, e.g. neuroectodermal cell types. Thereby, a lot of protocols were reported for differentiating hMSCs into specific neuron cells. Within those induction methods, most of them emphasized that the key for controlling the differentiating procedure is the addition of growth factors, cytokines, and even chemicals. In this study, we directed hMSCs to neural differentiation by applying an alkaline medium supplemented with 2% fetal bovine serum (FBS). Results from RT-PCR disclosed that some neuron-specific genes including NeuroD were exclusively expressed in the differentiated cells. Besides cell morphological change, changes of proteome profiling by neural induction was observed with the conventional proteomic method using two-dimensional gel electrophoresis (2-DE) for separating proteins and mass spectrometry for the identification of separated proteins. The 2-DE was also used to profile the proteins secreted by hMSCs and differentiated cells. Differentially expression of intracellular and extracellular proteomes between hMSCs and differentiated cells was evident. For example, the Alzheimer’s disease related mitochondrial antioxidant enzyme MnSOD increased about 21 times in the differentiated cells, and other proteins like MAP6 protein with the ability to repair synapses and IGFBP-7 protein which can regulate PI3K/Akt signaling pathway also existed after the induction procedure. In addition, hMSCs modified with targeting DNA hypermethylation of genes in Salvador/Warts/Hippo (SWH) pathway were also studied for the neural differentiation. Although the SWH knockout hMSCs were found to have a higher rate of expansion and differentiation, their neural differentiation pattern was very similar to that of intact hMSCs.