K.D. Bozov1, A.N. Velikanov2, V.S. Usatova3, M.A. Berestovoy4, M.E. Illarionova5, L.S. Gonzhal6, S.S. Dzhauari7, L.N. Shkarina8, A.L. Primak9, E.V. Semina10, L.M. Samokhodskaya11, P.S. Klimovich12, V.S. Popov13, E.A. Neyfeld14, M.N. Karagyaur15

1,2,5–15 Medical Research and Educational Institute Lomonosov Moscow State University (Moscow, Russia)

3,4 Federal State Budgetary Institution «Federal Center of Brain Research and Neurotechnologies» of the Federal Medical and Biological Agency (Moscow, Russia)

1 kir-bozov@yandex.ru, 2 av-bioem@mail.ru, 3 nikausatova@gmail.com, 4 berestovoy@fccps.ru, 5 mar729i63illar90@yandex.ru, 6 gonzhallev@yandex.ru, 7 stalik.djauari@yandex.ru, 8 shliliyan@mail.ru, 9 primak.msu@mail.ru, 10 e-tal@yandex.ru, 11 samokhodskay@gmail.com, 12 lex2050@mail.ru, 13 galiantus@gmail.com, 14 ea.neyfeld@mail.ru, 15 m.karagyaur@mail.ru

Lentiviral transduction of induced pluripotent stem cells (iPSCs) and neural progenitor cells (NPCs) is an important but challenging task that is often required in a wide range of research applications. Lentiviral vectors are successfully used for the transduction of multipotent mesenchymal stromal cells, but iPSCs and NPCs are very sensitive to such modifications. For this reason, increasing the efficiency of their modification without significantly altering the viability of iPSC and NPC cultures by optimizing the lentiviral particle transduction procedure is a relevant experimental task.

The aim of the study was to investigate the efficiency of various protocols of transduction of induced pluripotent stem cells and human neural progenitor cells with a model lentiviral vector carrying the red fluorescent protein gene.

The optimal ratio of efficacy and toxicity for iPSC cultures was achieved using 4-fold concentrated lentiviral particle preparation (MOI 56) during 18-hour incubation. In NPC cultures, the most effective and least toxic was the regimen of 4-hour incubation with non-concentrated 1x lentiviral particle preparation (MOI 14).

The data obtained allows one to select the optimal parameters for lentiviral transduction of iPSCs and NPSCs,that facilitates creating cell models of human diseases, enables the study of the functional and clinical significance of certain genomic or biochemical disorders, and promotes the development of promising therapeutic approaches.

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