Mesenchymal Stem Cells Engineering: Microcapsules-Assisted Gene Transfection and Magnetic Cell Separation

Albert R. Muslimov, Alexander S. Timin, Aleksandra V. Petrova, Olga S. Epifanovskaya, Alena I. Shakirova, Kirill V. Lepik, Andrey Gorshkov, Eugenia V. Il'Inskaja, Andrey V. Vasin, Boris V. Afanasyev, Boris Fehse, Gleb B. Sukhorukov

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

Stem cell engineering - the manipulation and functionalization of stem cells involving genetic modification - can significantly expand their applicability for cell therapy in humans. Toward this aim, reliable, standardized, and cost-effective methods for cell manipulation are required. Here we explore the potential of magnetic multilayer capsules to serve as a universal platform for nonviral gene transfer, stem cell magnetization, and magnetic cell separation to improve gene transfer efficiency. In particular, the following experiments were performed: (i) a study of the process of internalization of magnetic capsules into stem cells, including capsule co-localization with established markers of endo-lysosomal pathway; (ii) characterization and quantification of capsule uptake with confocal microscopy, electron microscopy, and flow cytometry; (iii) intracellular delivery of messenger RNA and separation of gene-modified cells by magnetic cell sorting (MACS); and (iv) analysis of the influence of capsules on cell proliferation potential. Importantly, based on the internalization of magnetic capsules, transfected cells became susceptible to external magnetic fields, which made it easy to enrich gene-modified cells using MACS (purity ∼95%), and also to influence their migration behavior. In summary, our results underline the high potential of magnetic capsules in stem cell functionalization, namely (i) to increase gene-transfer efficiency and (ii) to facilitate enrichment and targeting of transfected cells. Finally, we did not observe a negative impact of the capsules used on the proliferative capacity of stem cells, proving their high biocompatibility.

Original languageEnglish
Pages (from-to)2314-2324
Number of pages11
JournalACS Biomaterials Science and Engineering
Volume3
Issue number10
DOIs
Publication statusPublished - 9 Oct 2017

Keywords

  • cell magnetization
  • gene delivery
  • in vitro cell separation
  • internalization pathway
  • magnetic capsules
  • mesenchymal stem cells

ASJC Scopus subject areas

  • Biomaterials
  • Biomedical Engineering

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