| 刘天庆,戴明舒,葛丹,李香琴,马学虎,崔占峰.神经球内神经干/祖细胞活性与代谢研究[J].,2008,(6):811-818 |
| 神经球内神经干/祖细胞活性与代谢研究 |
| Study of viability and metabolism parameters of NSPCs |
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| DOI：10.7511/dllgxb200806006 |
| 中文关键词: 神经干细胞 神经球 代谢 传质 |
| 英文关键词: neural stem cells(NSCs) neurosphere metabolism mass transfer |
| 基金项目:基金项目 [HTSS]国家自然科学基金资助项目(30670525). |
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| 摘要点击次数: 1566 |
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| 中文摘要: |
| 为定量研究静态无血清悬浮培养的神经球内神经干/祖细胞的活性及代谢情况，取孕14 d的小鼠胚胎，从海马脑组织分离出神经干/祖细胞，培养过程中跟踪测定神经球直径的分布变化，利用CCK-8法测定活细胞数，同时对培养上清进行葡萄糖、乳酸、谷氨酰胺和氨的定量测定，并对相关细胞代谢动力学参数进行计算和分析．结果表明在培养过程中，神经球内细胞活性及细胞代谢均随着球直径的增加而减弱．在神经球平均直径为100 μm左右，培养基中葡萄糖、谷氨酰胺浓度分别为36.38 mmol/L和1.33 mmol/L时，球心细胞生长开始受到抑制；而当神经球直径主体分布于100~150 μm，培养基中葡萄糖浓度为31.11 mmol/L、谷氨酰胺浓度为1.15 mmol/L时，神经球内部细胞死亡数开始大于增殖数，同时细胞代谢降低到很低水平．表明随着神经球直径的增加而造成的传质困难对于其内部细胞的活性和代谢有重要影响． |
| 英文摘要: |
| The aim of this study is to understand how the metabolism and viability of the cells in neurospheres changed with the increase of neurospheres′ diameters, the NSPCs (neural stem progenitor cells ) were dissociated from the hippocampus tissue of embryonic 14 d (E14) mouse. During the culture, the diameter distribution of neurospheres and the cell viability were detected. Meanwhile, the concentrations of glucose, lactic acid, glutamine and ammonium in the media were measured, and the corresponding cellular metabolic rates were calculated and analyzed. The results show that the cell viability decreases with the increase of neurospheres′ diameters. When the diameter reaches about 100 μm, and the concentrations of glucose and glutamine are 36.38 mmol/L and 1.33 mmol/L, the growing of central cells in neurospheres begins to be inhibited. Furthermore, when the diameter reaches about 100-150 μm, and the concentrations of glucose and glutamine are 31.11 mmol/L and 1.15 mmol/L, the number of dead cells inside neurospheres exceeds that of proliferative cells. And the metabolism of the cells declines to very low level. In conclusion, the mass transfer limitation caused by the increase of neurospheres′ diameters has serious impact on the cell viability and metabolism. |
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