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通过流式细胞术分析培养的神经母细胞瘤细胞中的聚乙二醇(PEG)融合:技术与优化

Analysis of polyethylene glycol (PEG) fusion in cultured neuroblastoma cells via flow cytometry: Techniques & optimization.

作者信息

Hoffman Ashley N, Bamba Ravinder, Pollins Alonda C, Thayer Wesley P

机构信息

Vanderbilt School of Medicine, 1161 21st Ave S # T1217, Nashville, TN 37232, United States.

Department of Plastic Surgery, Vanderbilt University Medical Center, 1161 21st Ave South, MCN S-2221, Nashville, TN, United States.

出版信息

J Clin Neurosci. 2017 Feb;36:125-128. doi: 10.1016/j.jocn.2016.10.032. Epub 2016 Nov 4.

Abstract

Polyethylene glycol (PEG) has long been used as a membrane fusogen, but recently it has been adopted as a technique for peripheral nerve repair. Vertebrate models using PEG fusion have shown improved outcomes when PEG is applied during repair of severed peripheral nerves. The cellular mechanism of PEG fusion in the peripheral nerve repair model has not previously been assessed via flow cytometry. PEG fusion was assessed in this experiment by dying B35 rat neuroblastoma cells with different color fluorescent labels. The different color cells were combined and PEG was applied in concentrations of 50%, 75% and 100%. The amount of cell fusion was assessed via flow cytometry as the percentage of double positive cells. Results showed increasing fusion and decreasing viability with increasing concentrations of PEG.

摘要

聚乙二醇(PEG)长期以来一直被用作膜融合剂,但最近它已被用作一种周围神经修复技术。使用PEG融合的脊椎动物模型表明,在修复切断的周围神经时应用PEG可改善结果。此前尚未通过流式细胞术评估周围神经修复模型中PEG融合的细胞机制。在本实验中,通过用不同颜色的荧光标记物对B35大鼠神经母细胞瘤细胞进行染色来评估PEG融合。将不同颜色的细胞合并,并以50%、75%和100%的浓度应用PEG。通过流式细胞术将细胞融合量评估为双阳性细胞的百分比。结果表明,随着PEG浓度的增加,融合增加,活力降低。

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