The Bio-effect of Polyelectrolyte Multilayer Films on Neural Stem/Progenitor Cells

• Main Authors: Yu Chieh Wu, 吳渝婕
• Other Authors: I. C. Lee
• Format: Others
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/24809695933861986115

碩士 === 長庚大學 === 生化與生醫工程研究所 === 101 === The mammalian central nervous system disorders, such as stroke, Parkinson’s disease, Multiple Sclerosis, spinal cord injury, has little capacity for self-repair after injury, and neurons are not able for proliferation. Neural stem/progenitor cells (NSPCs) have the ability of self-renewal, proliferation and differentiation. Previous studies have showed that the regulation of stem cell microenviroment could promote the growth and differentiation of NSPCs. Fabrication of a proper microenvironment for NSPCs on proliferation or specific differentiation lineage will give hopes on the application of neural repair and regeneration. In this study, supported lipid bilayer (SLB) was utilized to build a biomimetic system, and poly-L-lysine and poly-L-glutamic acid were chosen to assemble sequential layer-by-layer structure and to conjugate with SLB to form a series of SLB-PEM films. NSPCs were cultured on these substrates, and the effects of differentiation were observed. In addition, in comparison of the bioeffects of PEM films system and SLB-PEM films system were also investigated. Image J software was used to quantify the process length and immunestaining method was used to observe the differentiated cells and synapse function (MAP2 for neuron, GFAP for astrocyte, DAPI for nuclei, Synapsin I for snapsin), FM1-43 lipid dye was used to test the activity of synapse vesicle and western blot was used to quantify the protein expression of differentiated cells. Our results showed that NSPCs could attach and differentiate fast no matter on PEM films or SLB-PEM films after 2 days culture and the process outgrowth were highly promoted, especially on PLL terminating layer, processes could build good neural network. In addition, high percentage neurons with functional synapse were generated as the increase of layer number in both PEM and SLB-PEM culture systems. This result was more obvious to see under the niches of SLB based films and the percentage of neuron differentiation could reach to 80%. Western blot analysis also revealed that large amount of functional neurons were generated as the layer number increased in SLB-PEM system. In this study, a serum and chemical agent -free biomimetic system was built by using SLB and nature polypeptide materials, which could promote the generation of large amount of functional neurons and build a good neural network with long neurite outgrowth, and this result was very helpful at the application of neuron damage repair.