CAJ | 학술논문

目的:探讨外源性一氧化氮(NO)对人胚胎神经干细胞(NSCs)增殖的抑制作用.方法:取4月龄(±15d)正常孕妇引产胎儿大脑皮质组织分离代NSCs,免疫荧光化学法鉴定.培养基中加入不同浓度的硝普钠后,Griess还原法检测培养液中NO氧化产物的含量;MTT法测定细胞活力.结果:加入不同浓度硝普钠24h后,培养液中NO氧化产物的含量随硝普钠浓度的增高而增加,且显著高于对照组.MTT法显示经硝普钠作用后的NSCs活细胞数较对照组明显减少,并呈浓度相关性;在去除硝普钠后继续培养的NSCs仍可保持增殖能力.结论:外源性NO对培养状态下的人胚胎NSCs增殖有抑制作用.
목적:탐토외원성일양화담(NO)대인배태신경간세포(NSCs)증식적억제작용.방법:취4월령(±15d)정상잉부인산태인대뇌피질조직분리대NSCs,면역형광화학법감정.배양기중가입불동농도적초보납후,Griess환원법검측배양액중NO양화산물적함량;MTT법측정세포활력.결과:가입불동농도초보납24h후,배양액중NO양화산물적함량수초보납농도적증고이증가,차현저고우대조조.MTT법현시경초보납작용후적NSCs활세포수교대조조명현감소,병정농도상관성;재거제초보납후계속배양적NSCs잉가보지증식능력.결론:외원성NO대배양상태하적인배태NSCs증식유억제작용.
Objective: To study the inhibitive effect of exogenous nitric oxide (NO) on the proliferation of human fetal neural stem cells (NSCs). Methods: The primary NSCs from the cerebral cortex of 4-month old human fetuses were cultured in the serum-free medium,and identified with indirect immunoflurenscence cytochemistry. On day 5, the NSCs were cultured in the medium supplemented with different concentrations of sodium nitroprusside (SNP), and 24h later the NO content in the medium was measured with the Griess assay.Then half of the cultured cells were measured with MTT assay to detect the amount of living cells, and the remaining cells were kept in culture medium without SNP for another 24h, followed by MTT assay. Results: The NO content in the experimental groups was increased with the SNP concentration, and significantly higher than that in the control group. The amount of living cells in the experimental groups was significantly decreased in a concentration-dependent manner compared with that in the control group. The NSCs still retained their ability to proliferate when SNP had been removed. Conclusion: Exogenous NO can inhibit the proliferation of NSCs from human fetus in vitro.