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在以微流体技术加工的聚乙烯亚胺(polyethyleneimine,PEI)及层粘连蛋白(laminin,LN)混合物材料上,培养纹状体神经元图案式黏附、生长,为制备神经芯片打好基础。以微流体印刷方法在硅片上微加工四种不同的黏附底物:LN、带正电荷的多聚赖氨酸(poly-L-lysine,PLL)、PEI和PEI+LN混合物。体外分离培养新生乳鼠纹状体神经元,评价神经元在不同图案化黏附底物上的黏附存活、突起生长状态及形成流道图案的差异;在PEI+LN混合物图案中观察不同线间距对构建神经元流道图案的效果。结果表明,在PEI、PLL及PEI+LN图案表面生长的神经元数量明显大于LN组;神经元在PEI+LN混合底物上与单纯PEI、LN、PLL相比更能形成完整的流道图案,在流道间距处于150μm及200μm时更为清晰。研究证实,通过微流体微加工后,由PEI与LN混合形成的粘附底物经优化流道图案规格后,能有效地构建神经元流道图案,有利于制备神经芯片。
The cultured neurons of striatum were adhered and grew patternally on the surfaces of polyethyleneimine (PEI) and laminin (LN) mixture treated by microfluid technology, which laid the foundation for the preparation of neural microchip. Four different adhesion substrates, LN, positively charged poly-L-lysine (PLL), PEI and PEI + LN mixtures were microfabricated on silicon wafers by microfluidic printing. The neurons of neonatal rat striatum were isolated and cultured in vitro. The adhesion survival, the growth state of the neurons and the pattern of forming the flow channel on different patterned adhesion substrates were evaluated. In the PEI + LN mixture pattern, The effect of constructing neuronal runner patterns. The results showed that the number of neurons grown on the surface of PEI, PLL and PEI + LN patterns was significantly larger than that of LN group. Neurons formed a complete flow pattern on PEI + LN mixed substrate compared with pure PEI, LN and PLL , The runner spacing at 150μm and 200μm when clearer. The study confirmed that after the micro-fluid micro-machining, the adhesion substrate formed by mixing PEI with LN optimized the flow pattern specifications, which can effectively construct the neuronal flow channel pattern and facilitate the preparation of the neural chip.