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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,h PSC)向神经元的定向分化。方法:应用25.0、12.5、8.5、5.0μg/m L 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h PSC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(25.0μg/m L)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(5.0μg/m L)消除支原体所需给药时间较长,12.5μg/m L浓度组对细胞生长有不利影响,而8.5μg/m L浓度组对细胞生长影响较小。h PSC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8.5μg/m L的Plasmocin加药1周即可彻底消除h PSC中支原体的感染,并且不会对其定向分化能力造成影响。
OBJECTIVE: To explore the optimal concentration and the number of days required for Plasmocin to eradicate mycoplasma infection of human pluripotent stem cells, and to detect the directional differentiation of human pluripotent stem cell (h PSC) into neurons after mycoplasma infection. Methods: Mycoplasma pneumoniae was treated with Plasmocin at different concentrations of 25.0, 12.5, 8.5 and 5.0 μg / mL, and the results were compared by PCR. The h PSC with mycoplasma hominis was cultured and differentiated. Results: Plasmocin at four concentrations could eliminate mycoplasma infection. However, cytotoxic reaction occurred in the high concentration group (25.0 μg / mL), but fewer mycoplasma survived after the elimination of Mycoplasma. In the low concentration group (5.0 μg / m L) administration of Mycoplasma for a long time, 12.5μg / m L concentration group had an adverse effect on cell growth, while 8.5μg / m L concentration group had little effect on cell growth. h PSC efficiently differentiated into neurons in the forebrain after elimination of mycoplasma infection, and no mycoplasma was detected after 9 months. Conclusion: The application of 8.5μg / m L Plasmocin for 1 week can completely eliminate the mycoplasma infection in h PSC, and will not affect the ability of its directional differentiation.