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[影响用磁性纳米颗粒标记NIH 3T3细胞的因素]

[Factors Affecting the Labeling of NIH 3T3 Cells with Magnetic Nanoparticles].

作者信息

Kandarakov O F, Demin A M, Popenko V I, Leonova O G, Kopantseva E E, Krasnov V P, Belyavsky A V

机构信息

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia.

Postovsky Institute of Organic Synthesis, Ural Branch, Russian Academy of Sciences, Yekaterinburg, 620990 Russia.

出版信息

Mol Biol (Mosk). 2020 Jan-Feb;54(1):114-127. doi: 10.31857/S0026898420010073.

Abstract

The factors that affect the labeling of NIH 3T3 murine fibroblasts with Fe3O4-based magnetic nanoparticles (MNPs) were studied using MNPs produced by the gas condensation and solution precipitation methods and MNPs surface-modified with 3-aminopropylsilane or L-lysine. The production method, surface modifications, the particle concentration and size, the state of the cell population, and the method of MNP introduction were found to substantially affect the efficiency of MNP binding by cells. In particular, large MNP clusters may occur in MNP suspensions in DMSO, and their disruption by sonication increased the percent yield of magnetically labeled cells. Static incubation of a cell suspension led to a more efficient labeling as compared with continuous agitation. Cells attached to a plastic support could be labeled to a higher degree than cells in suspension, but required substantially longer incubations with MNPs. MNP centrifugation on cell layers (magnetic spinoculation) significantly increased the rate and efficiency of labeling. The stability of magnetic labeling was shown to depend on the MNP dose during labeling. Electron microscopy studies demonstrated that MNPs were associated with the cell surface after 20-min incubation with cells and were mostly in the cell interior after 4-h incubation. The results of the study may be useful for preparation and application of magnetized cell samples.

摘要

使用通过气体冷凝法和溶液沉淀法制备的基于Fe3O4的磁性纳米颗粒(MNP)以及用3-氨丙基硅烷或L-赖氨酸进行表面修饰的MNP,研究了影响NIH 3T3小鼠成纤维细胞用MNP标记的因素。发现生产方法、表面修饰、颗粒浓度和尺寸、细胞群体状态以及MNP引入方法对细胞结合MNP的效率有显著影响。特别是,在二甲基亚砜(DMSO)中的MNP悬浮液中可能会出现大的MNP簇,通过超声处理破坏它们可提高磁性标记细胞的百分产率。与持续搅拌相比,细胞悬浮液的静态孵育导致更有效的标记。附着在塑料支持物上的细胞比悬浮细胞能被标记到更高的程度,但与MNP孵育所需的时间要长得多。在细胞层上进行MNP离心(磁性旋转接种)显著提高了标记的速率和效率。结果表明,磁性标记的稳定性取决于标记过程中的MNP剂量。电子显微镜研究表明,与细胞孵育20分钟后,MNP与细胞表面相关,孵育4小时后大多位于细胞内部。该研究结果可能对磁化细胞样品的制备和应用有用。

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