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背景:星形细胞对神经元有提供营养、支持及调节突触活性作用,但它对神经元发育的影响还尚不清楚。目的:探讨体外培养的SpragueDawley大鼠大脑皮质星形细胞对PC12神经元突起生长发育的作用。设计:完全随机设计,对照实验研究。方法:以培养的星形细胞与PC12神经元按不同细胞数目比例(50∶1～1∶1)共同培养,并用其制备的条件培养液培养PC12细胞。主要观察指标:用快速灵敏的MTT比色法测定PC12神经元的细胞活力,用光学相差显微镜观察PC12细胞形态学变化。结果:①星形细胞条件培养液可增强PC12细胞活力(MTT测定的A值由0.255±0.012提高到0.510±0.036,P<0.001),却不能促使PC12神经元突起的生出。②当将星形细胞与PC12细胞按30∶1～1∶1的比例共同培养时,既可提高PC12细胞折光性和光晕又可促使其突起的生长;但按50∶1～40∶1的比例共同培养时,只观察到提高PC12细胞折光性和光晕,而无促使其突起生长发育的作用。结论:PC12神经元细胞活力的提高与星形细胞分泌到条件培养液中的可溶性因子有关,而PC12神经元突起生长发育可能是和与星形细胞的直接接触以及二者的细胞数目比有关。
BACKGROUND: Astrocytes provide nutrients to neurons, support and regulate synaptic activity, but its effect on neuronal development remains unclear. Objective: To investigate the effect of cultured cerebral cortex astrocytes on the growth and development of PC12 neurons in Sprague Dawley rats. Design: Completely randomized design, controlled experimental study. Methods: Cultured astrocytes and PC12 neurons were co-cultured at a ratio of different cells (50: 1 ~ 1: 1). PC12 cells were cultured in the conditioned medium. MAIN OUTCOME MEASURES: Cell viability of PC12 neurons was measured by MTT colorimetric assay. Morphological changes of PC12 cells were observed by optical phase contrast microscope. RESULTS: ① Astrocyte conditioned medium enhanced the viability of PC12 cells (MTT assay A value increased from 0.255 ± 0.012 to 0.510 ± 0.036, P <0.001), but did not promote the neurite outgrowth in PC12 cells. ② When astrocytes and PC12 cells are co-cultured in a ratio of 30: 1 to 1: 1, the PC12 cells can improve the refraction and the halo of the PC12 cells and promote the growth of the protrusions. However, the ratio of 50: 1 to 40: 1 Proportional co-culture, only to improve the PC12 cell refraction and halo, without promoting its role in the process of protruding growth and development. CONCLUSION: The increased viability of PC12 neurons is related to the soluble factor secreted by astrocytes into conditioned media. The growth and development of PC12 neurons may be related to the direct contact with astrocytes and the cell number ratio.