Plasmocin对人多能干细胞支原体感染的消除研究
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国家自然科学基金面上项目(81471301);江苏省双创人才基金(KY216R201510);南京医科大学引进人才启动基金(KY109RC011305)


Application of Plasmocin in eliminating mycoplasma infection of human pluripotent stem cell
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    摘要:

    目的:探究应用Plasmocin消除人多能干细胞支原体感染时的最佳浓度和所需天数,检测消除支原体感染后的人多能干细胞(human pluripotent stem cell,hPSC)向神经元的定向分化-方法:应用25.0-12.5-8.5-5.0 μg/mL 4种不同浓度的Plasmocin进行支原体感染的消除,用PCR比较消除结果;对已消除支原体的hPSC进行维持培养以及定向分化-结果:应用4种浓度的Plasmocin均能消除支原体的感染,但其中高浓度组(25.0 μg/mL)出现细胞毒性反应,消除支原体后存活下来细胞较少,而低浓度组(5.0 μg/mL)消除支原体所需给药时间较长,12.5 μg/mL浓度组对细胞生长有不利影响,而8.5 μg/mL浓度组对细胞生长影响较小-hPSC在消除支原体感染后可高效分化至前脑神经元,并且9个月后未检测出支原体-结论:应用8.5 μg/mL的Plasmocin加药1周即可彻底消除hPSC中支原体的感染,并且不会对其定向分化能力造成影响-

    Abstract:

    Objective:To investigate the most effective dosage and optimal time window of Plasmocin in eliminating mycoplasma infection,and detect whether the drug exert negative effects on human pluriotent stem cell(hPSC) culture. Methods:hPSCS were treated with 25.0,12.5,8.5,5.0 μg/mL of Plasmocin. The efficiency for removing mycoplasma infection was compared by PCR. Directly differentiation of hPSC to forebrain neurons after rescued. Results:All of the concentrations for Plasmocin eradicated mycoplasma infection. However,the highest dosage (25.0 μg/mL)lead to toxic effects and the lowest dosage(5.0 μg/mL)required longer treatment time. The 12.5 μg/mL group had adverse effects on cell growth but the 8.5 μg/mL group contributed to hPSCs maintenance. After mycoplasma elimination,hPSC efficiently differentiated to forebrain neurons. Furthermore,after 9 months post-Plasmocin treatment,mycoplasam was undetected on hPSC. Conclusion:One week treatment with 8.5 μg/mL Plasmocin completely eradicated mycoplasma contamination without affecting hPSC maintenance and differentiation.

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曹诗颖,瞿庄茵,袁 方,刘 妍. Plasmocin对人多能干细胞支原体感染的消除研究[J].南京医科大学学报(自然科学版),2016,(8):905-911

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  • 收稿日期:2016-03-17
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  • 在线发布日期: 2016-08-19
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