目的：体外观察磁刺激对原代神经元迁移的影响。方法原代培养 SD 大鼠皮质神经元，分为正常组（control，C 组）、假刺激组（shame，S 组）、40％最大输出强度组（40％ of maximum intensity of stimulation，0．4T 组）、60％最大输出...目的：体外观察磁刺激对原代神经元迁移的影响。方法原代培养 SD 大鼠皮质神经元，分为正常组（control，C 组）、假刺激组（shame，S 组）、40％最大输出强度组（40％ of maximum intensity of stimulation，0．4T 组）、60％最大输出强度组（60％ of maximum intensity of stimulation，0．6T 组）。各组细胞接种于 transwell 小室内，自细胞接种后第2～6天接受磁刺激，连续刺激5 d，各组细胞于第6天同一时间染色，将 transwell 小室倒置，光学显微镜下计数。结果C 组有（2．6±0．548）个、S 组有（3．0±0．707）个细胞迁移，2组比较差异无统计学意义（P ＞0．05）；0．4T 组有（17．40±1．673）个、0．6T 组有（18．40±1．572）个细胞迁移，迁移的数量较 C 组及 S 组明显增多（P ＜0．05）。结论磁刺激可促进体外原代神经元迁移。展开更多
High-throughput quantitative approaches to study axon growth behaviors have remained a challenge.We have developed a 1024-chamber microfluidic gradient generator array that enables large-scale investigations of axon g...High-throughput quantitative approaches to study axon growth behaviors have remained a challenge.We have developed a 1024-chamber microfluidic gradient generator array that enables large-scale investigations of axon guidance and growth dynamics from individual primary mammalian neurons,which are exposed to gradients of diffusible molecules.Our microfluidic method(a)generates statistically rich data sets,(b)produces a stable,reproducible gradient with negligible shear stresses on the culture surface,(c)is amenable to the long-term culture of primary neurons without any unconventional protocol,and(d)eliminates the confounding influence of cell-secreted factors.Using this platform,we demonstrate that hippocampal axon guidance in response to a netrin-1 gradient is concentration-dependent—attractive at higher concentrations and repulsive at lower concentrations.We also show that the turning of the growth cone depends on the angle of incidence of the gradient.Our study highlights the potential of microfluidic devices in producing large amounts of data from morphogen and chemokine gradients that play essential roles not only in axonal navigation but also in stem cell differentiation,cell migration,and immune response.展开更多
文摘目的：体外观察磁刺激对原代神经元迁移的影响。方法原代培养 SD 大鼠皮质神经元，分为正常组（control，C 组）、假刺激组（shame，S 组）、40％最大输出强度组（40％ of maximum intensity of stimulation，0．4T 组）、60％最大输出强度组（60％ of maximum intensity of stimulation，0．6T 组）。各组细胞接种于 transwell 小室内，自细胞接种后第2～6天接受磁刺激，连续刺激5 d，各组细胞于第6天同一时间染色，将 transwell 小室倒置，光学显微镜下计数。结果C 组有（2．6±0．548）个、S 组有（3．0±0．707）个细胞迁移，2组比较差异无统计学意义（P ＞0．05）；0．4T 组有（17．40±1．673）个、0．6T 组有（18．40±1．572）个细胞迁移，迁移的数量较 C 组及 S 组明显增多（P ＜0．05）。结论磁刺激可促进体外原代神经元迁移。
基金This work was supported by a grant from the National Institutes of Health(1R01NS064387).
文摘High-throughput quantitative approaches to study axon growth behaviors have remained a challenge.We have developed a 1024-chamber microfluidic gradient generator array that enables large-scale investigations of axon guidance and growth dynamics from individual primary mammalian neurons,which are exposed to gradients of diffusible molecules.Our microfluidic method(a)generates statistically rich data sets,(b)produces a stable,reproducible gradient with negligible shear stresses on the culture surface,(c)is amenable to the long-term culture of primary neurons without any unconventional protocol,and(d)eliminates the confounding influence of cell-secreted factors.Using this platform,we demonstrate that hippocampal axon guidance in response to a netrin-1 gradient is concentration-dependent—attractive at higher concentrations and repulsive at lower concentrations.We also show that the turning of the growth cone depends on the angle of incidence of the gradient.Our study highlights the potential of microfluidic devices in producing large amounts of data from morphogen and chemokine gradients that play essential roles not only in axonal navigation but also in stem cell differentiation,cell migration,and immune response.
