Mesenchymal stem cells engineering: microcapsules assisted gene transfection and magnetic cell separation.

TY - JOUR

T1 - Mesenchymal stem cells engineering: microcapsules assisted gene transfection and magnetic cell separation.

AU - Muslimov, Albert R

AU - Timin, Alexander S

AU - Petrova , AV

AU - Epifanovskaya, Olga S

AU - Shakirova, Alena I.

AU - Lepik, Kirill V.

AU - Gorshkov , A

AU - Il’inskaja , EV

AU - Vasin, Andrey V.

AU - Afanasyev, Boris V.

AU - Fehse, Boris

AU - Sukhorukov, Gleb B

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

M3 - SCORING: Journal article

VL - 3

SP - 2314

EP - 2324

JO - ACS BIOMATER SCI ENG

JF - ACS BIOMATER SCI ENG

SN - 2373-9878

IS - 10

ER -