Development of GaN chips to culture cerebellar granule neurons，neural stem/precursor cells and PC12 cells
Date Issued
2009
Date
2009
Author(s)
Chen, Chi-Ruei
Abstract
Neurons can only grow well in body by means of neurotrophic factors and afferent neuron stimulus. Cerebellar granule neurons belong to highly uniform neural culture model, and have been widely used in neuron growth and apoptosis mechanism researches. Neural stem cells are functional cells with multi-differentiation potency, having potential of repairing central nerve. Cell culture substrate for in vitro cultivation has always been a major factor in cell culture. This study examined cell adhesion and differentiation difference when adopting bioactive molecule-polylysine, silicon chip, GaN, GaAs and commonly used cell culture dish (polystyrene) as substrate of cultivating neurons or neural stem cells respectively. In addition, to understand the cause of such differences, this study selected neuronal pheochromocytoma strain was selected in to analyze signal transduction pathway, and cell behavior variation can be attributed to protein expression difference. hapter 1 introduces application and comparison of central neuron, neural stem cell, chromaffin neuron and semiconductor material used on cell culture. hapter 2 discusses neuron adhesion and differentiation after cultured on GaN, silicon substrate and tissue culture polystyrene (TCPS) for 3-6 days. As shown by optical micrographs, after cultured for three days, most neurons could form cell adhesion and differentiation on every above substrate. And after cultured for six days, neurons on GaN substrate showed more differentiation, neurons on silicon substrate reported deteriorated adhesion and differentiation. Furthermore, cell adhesion degree inspection on lactate dehydrogenase showed the same trend. hapter 3 describes the long-term synapsis performance of GaN cultured neuron, showing that, GaN has better retention of synapsis function, as compared to polylysine and TCPS. Relative cell death test also showed that GaN could better promote cell survival, probably due to phosphorylation of Akt on GaN substrate in earlier time. hapter 4 examines neural stem cell differentiation on GaN and polylysine respectively. Cultivation showed that both substrates could enable cell adhesion and differentiation, GaN was more likely than polylysine to enable neural stem cell differentiation along neuron path, probably due to inhibited activity of glycogen synthase kinase-3β (GSK-3β). On the other hand, GaN is capable of promoting long term adhesion of neural stem cell and cell survival. hapter 5 takes chromaffin cell as the model of research on neuron signal transduction pathway. The experiment found that, GaN may suppress neuron apoptosis via Akt/GSK-3β/caspase-3 path, and thus promote neuron survival; lactate dehydrogenase test indicated the same trend.hapter 6 concludes the contribution of this study that Lewis acid-base property derived from Group III/V semiconductor GaN single crystal could promote adhesion and differentiation of neurons and neural stem cells.
Subjects
neuron
neural stem cell
PC12
GaN
synapsis
adhesion
differentiation
Type
thesis
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