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

  • MOLECULAR CELL BIOLOGY
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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.

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Funding

Experiments illustrated with Figs. 1‒10 were supported by the Russian Science Foundation (project no. 18-14-00300). Synthesis and physicochemical characterization of MNPs used in this work (Figs. S1‒S3) were supported by a state contract (project no. AAAA-A18-118020290116-5s) and were carried out at the Collective Access Center for Spectroscopy and Analysis of Organic Compounds (SAOS Center).

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Authors and Affiliations

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

    O. F. Kandarakov, V. I. Popenko, E. E. Kopantseva & A. V. Belyavsky

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

    A. M. Demin, O. G. Leonova & V. P. Krasnov

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  1. O. F. Kandarakov
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  2. A. M. Demin
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  3. V. I. Popenko
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  4. O. G. Leonova
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  6. V. P. Krasnov
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  7. A. V. Belyavsky
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Correspondence to A. V. Belyavsky.

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Translated by T. Tkacheva

Abbreviations: DMSO, dimethyl sulfoxide; DMF, dimethylformamide; IR, infrared; MNP, magnetic nanoparticle; TGA, thermal gravimetric analysis; US, ultrasound; PBS, phosphate-buffered saline; APS, 3-aminopropylsilane; APTMS, (3-aminopropyl)trimethoxysilane; TBTU, O-(1H-benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; DIPEA, N,N-diisopropylethylamine.

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Kandarakov, O.F., Demin, A.M., Popenko, V.I. et al. Factors Affecting the Labeling of NIH 3T3 Cells with Magnetic Nanoparticles. Mol Biol 54, 99–110 (2020). https://doi.org/10.1134/S0026893320010070

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