Re-evaluation of Culture Condition of PC12 and SH-SY5Y Cells Based on Growth Rate and Amino Acid Consumption
Re-evaluation of Culture Condition of PC12 and SH-SY5Y Cells Based on Growth Rate and Amino Acid Consumption
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DOI：
10.21873/invivo.11174
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发表时间：
2017-11-01
期刊：
影响因子：
2.3
通讯作者：
Takeshima, Hiroshi
中科院分区：
文献类型：
--
作者：
Sakagami, Hiroshi;Suzuki, Ryuichiro;Takeshima, Hiroshi
背景/目的：以往的研究者大多使用不同类型的培养基培养神经细胞。为了优化培养条件，我们比较了在DMEM或DMEM: Ham's F-12(1:1)：非必需氨基酸中生长的两种常用神经元模型大鼠PC12和人SH-SY5Y的生长速度和氨基酸消耗，并分别添加10%胎牛血清（分别称为DMEM和Mix）。材料与方法：采用MTT法监测细胞生长。脱蛋白后，用氨基酸分析法定量测定培养基中的氨基酸含量。结果：即使在极低的细胞密度下接种PC12细胞在无包被的普通培养皿中，不添加其条件培养基，也可以获得有效的细胞附着。PC12和SH-SY5Y细胞在DMEM中的增殖密度略高于Mix。DMEM对谷氨酰胺和必需氨基酸的利用率提高了约2倍。淀粉样肽如A β(1-42)和A β(25-35)抑制了它们近50%的生长。结论：基于DMEM对细胞附着、细胞生长、氨基酸利用和淀粉样肽敏感性的有利作用，本研究表明DMEM在寻找神经保护物质的研究中是有用的。
Background/Aim: Most of the previous investigators have used various types of media for the culture of nerve cells. In order to optimize the culture conditions, we compared the growth rate and amino acid consumption by two popular neuron models, rat PC12 and human SH-SY5Y, grown in DMEM or DMEM: Ham's F-12 (1: 1): non-essential amino acids, supplemented with 10% fetal bovine serum (referred to DMEM and Mix, respectively). Materials and Methods: Cell growth was monitored by the MTT method. Amino acids in the culture medium were quantitated by amino acid analysis after deproteinization. Results: Efficient cell attachment could be achieved even if PC12 cells were inoculated at extreme lower cell density in a non-coated plain dish, without addition of its condition medium. Both PC12 and SH-SY5Y cells proliferated up to slightly higher cell density in DMEM than in Mix. Approximately 2-fold higher utilization rate of glutamine and essential amino acids was observed in DMEM. Amyloid peptides such as A beta(1-42) and A beta(25-35) suppressed their growth nearly by 50%. Conclusion: The present study suggests the usefulness of DMEM for the study of searching neuroprotective substances, based on its favorable effects on cell attachment, cell growth and amino acid utilization as well as amyloid peptide sensitivity.