ssmuБюллетень сибирской медициныBulletin of Siberian Medicine1682-03631819-3684Siberian State Medical University, the Ministry of Healthcare of the Russian Federation10.20538/1682-0363-2020-4-143-150ssmu-4162Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL PAPERSМодификация макрофагов и моноцитов человека магнитными наночастицами in vitro для доставки, опосредованной клеткамиModification of human monocytes and macrophages by magnetic nanoparticles in vitro for cell-based deliveryhttps://orcid.org/0000-0001-9310-3196ПерекучаН. А.PerekuchaN. A.

мл. науч. сотрудник, Центр биологических исследований и биоинженерии Центральнойнаучно-исследовательской лаборатории

Россия, 634050, г. Томск, Московский тракт, 2

2, Moscow Trakt, Tomsk, 634050, Russian Federation

https://orcid.org/0000-0001-9216-9572СмолинаП. А.SmolinaP. A.

студент

Россия, 634050, г. Томск, Московский тракт, 2

2, Moscow Trakt, Tomsk, 634050, Russian Federation

https://orcid.org/0000-0002-4132-2327ДёминА. М.DeminA. M.

канд. хим. наук, ст. науч. сотрудник, лаборатория асимметрического синтеза

Россия, 620990, г. Екатеринбург, ул. С. Ковалевской, 22

22, S. Kovalevskoy Str., Yekaterinburg, 620990, Russian Federation

https://orcid.org/0000-0003-1411-6218КрасновВ. П.KrasnovV. P.

22, S. Kovalevskoy Str., Yekaterinburg, 620990, Russian Federation

https://orcid.org/0000-0001-7604-9722ПершинаА. Г.PershinaA. G.

канд. биол. наук, зав. центра биологических исследований и биоинженерии Центральной научно-исследовательской лаборатории; доцент, Исследовательская школа химических и биомедицинских технологий 

Россия, 634050, г. Томск, Московский тракт, 2

Россия, 634050, г. Томск, пр. Ленина, 30

2, Moscow Trakt, Tomsk, 634050, Russian Federation

30, Lenina Av., Tomsk, 634050, Russian Federation

allysyz@mail.ruСибирский государственный медицинский университет (СибГМУ)РоссияSiberian State Medical UniversityRussian FederationИнститут органического синтеза (ИОС) им. И.Я. Постовского Уральского отделения Российской академии наук (УрО РАН)РоссияPostovsky Institute of Organic Synthesis, Ural Branch of Russian Academy of SciencesRussian FederationСибирский государственный медицинский университет (СибГМУ); Национальный исследовательский Томский политехнический университет (НИ ТПУ)РоссияSiberian State Medical University; National Research Tomsk Polytechnic UniversityRussian Federation202007012021194143150Copyright © Перекуча Н.А., Смолина П.А., Дёмин А.М., Краснов В.П., Першина А.Г., 20212021Перекуча Н.А., Смолина П.А., Дёмин А.М., Краснов В.П., Першина А.Г.Perekucha N.A., Smolina P.A., Demin A.M., Krasnov V.P., Pershina A.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://bulletin.ssmu.ru/jour/article/view/4162

Цель исследования – разработать протокол модификации макрофагов и моноцитов человека магнитными наночастицами оксида железа (Fe3O4) in vitro.

Материалы и методы

Материалы и методы. Магнитные наночастицы оксида железа получены методом со-осаждения, покрыты силоксановой оболочкой и полиэтиленгликолем 3000. Макрофаги мыши линии RAW 264.7, моноциты периферической крови и макрофаги человека  инкубировали с магнитными наночастицами в течение 1–24 ч. Эффективность захвата наночастиц клетками оценивали феррозиновым методом и методом микроскопии с окрашиванием на железо по Перлсу. Исследование жизнеспособности клеток выполняли методом проточной цитофлуориметрии с использованием красителя SYTOX Green.

Результаты

Результаты. Инкубация макрофагов с магнитными наночастицами в концентрации ˃5 мкг/мл в течение 1 ч на ротаторе при 37 оС обеспечивает загрузку наночастиц в >99% клеток. Исследуемые магнитные наночастицы не оказывают негативных эффектов на жизнеспособность клеток. Клетки линии RAW 264.7, поглотившие наночастицы, сохраняют миграционную активность. Эффективность загрузки макрофагов магнитными наночастицами составляет ˃50 пкг (Fe)/клетку.

Заключение

Заключение. Макрофаги, загруженные магнитными наночастицами согласно предложенному протоколу, являются жизнеспособными, сохраняют способность к миграции и перспективны в качестве систем доставки, опосредованной клетками, для  диагностики и терапии опухоли.

The aim of the study was to develop a method for the modification of human monocytes/macrophages by iron oxide magnetic nanoparticles in vitro.

Materials and methods

Materials and methods. Iron oxide magnetic nanoparticles were obtained by a co-precipitation method and coated with a thin SiO2 layer and polyethylene glycol 3000. Murine macrophage-like cell line RAW 264.7, primary human monocytes and macrophages were incubated with magnetic nanoparticles for 1–24 hours. The efficiency of cellular uptake of nanoparticles was measured using a ferrozine-based method and microcopy with Perls’ Prussian blue staining. The cell viability was tested by fluorescent flow cytometry using SYTOX Green.

Results

Results. Incubation of RAW264.7 cell, human monocytes and macrophages with magnetic nanoparticles at a concentration ˃ 5 µg/mL on a rotator for 1 hour at 37 °С provides the loading of nanoparticles into > 99% of cells. The magnetic nanoparticles have no adverse effect on the cell viability. The RAW264.7 cells modified with nanoparticles showed no change in migration activity. The efficiency of the nanoparticle uptake by  macrophages was ˃50 pkg (Fe)/cell.

Conclusion

Conclusion. According to the proposed method, macrophages loaded with magnetic nanoparticles have proved viable, they retain the ability to migrate, and therefore can be used as cell-based delivery systems for tumor diagnostic and therapy.

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The authors declare that there are no conflicts of interest present.