Polybrene: Observations on cochlear hair cell necrosis and minimal lentiviral transduction of cochlear hair cells
Polybrene: Observations on cochlear hair cell necrosis and minimal lentiviral transduction of cochlear hair cells
复制标题
Polybrene：耳蜗毛细胞坏死和耳蜗毛细胞最小慢病毒转导的观察
DOI：
10.1016/j.neulet.2015.06.011
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发表时间：
2015-07-23
影响因子：
2.5
通讯作者：
He, Jingchun
中科院分区：
文献类型：
--
作者：
Han, Miaomiao;Yu, Dongzhen;He, Jingchun
聚苯乙烯被广泛用于增强病毒转导；然而，人们对其在增强耳蜗细胞慢病毒转导方面的作用知之甚少。在本研究中，我们研究了聚苯乙烯的细胞毒性作用，以及其对耳蜗细胞，特别是感觉毛细胞慢病毒转导的进一步影响。培养新生大鼠耳蜗基底膜，用0.1 ~ 10 μ g/mL聚苯乙烯处理24h，探讨耳蜗基底膜的耳毒性发展潜力。PI染色及TUNEL检测检测毛细胞坏死或凋亡情况。将不同剂量的慢病毒-绿色荧光蛋白加入到安全浓度的聚苯乙烯耳蜗器官型培养物中，孵育24小时，再培养48小时（在没有病毒和聚苯乙烯的情况下）。评估转导效率。结果表明，0.1 μ g/mL浓度的聚苯乙烯对耳蜗细胞是安全的，0.5 ~ 10 μ g/mL浓度的聚苯乙烯诱导毛细胞坏死呈剂量依赖性。然而，支持细胞没有受损。慢病毒载体转导到耳蜗细胞中，0.1 μ g/mL聚苯乙烯增强了转导效率。然而，无论是单独使用慢病毒载体还是在0.1 μ g/mL聚苯乙烯的存在下，毛细胞都很难被转导。聚苯乙烯对耳蜗毛细胞慢病毒转导的作用有待进一步研究。2015爱思唯尔爱尔兰有限公司版权所有。
Polybrene is widely used to enhance viral transduction; however, little is known about the utility thereof, in enhancing lentiviral transduction of cochlear cells. In the present study, we examined the cytotoxic effects of polybrene, and the further effects thereof, on lentiviral transduction of cochlear cells, especially sensory hair cells. Cochlear basilar membranes of newborn rats were cultured and treated with 0.1-10 mu g/mL polybrene for 24h to explore the potential development of ototoxicity. PI staining and TUNEL detection were used to evaluate necrosis or apoptosis of hair cell. Various doses of lentivirus-GFP were added to cochlear organotypic cultures with safe concentrations of polybrene, incubated for 24 h, and cultured (in the absence of the virus and polybrene) for a further 48 h. Transduction efficiencies were evaluated. The results showed that polybrene at 0.1 mu g/mL was safe to cochlear cells, and 0.5-10 mu g/mL concentration induced hair cell necrosis in a dose-dependent manner. However, supporting cells were not damaged. Lentiviral vectors transduced into cochlear cells and 0.1 mu g/mL polybrene enhanced transduction efficiency. However, hair cells were hardly transduced with lentiviral vectors either alone or in the presence of 0.1 mu g/mL polybrene. The use of polybrene to aid lentiviral transduction of cochlear hair cells requires further attention. (C) 2015 Elsevier Ireland Ltd. All rights reserved.