Application of the AAV-Syn-BDNF-EGFP Virus Vector as a Neuroprotective Agent in Modeling Hypoxia in vitro

The aim of the study was to develop the AAV-Syn-BDNF-EGFP virus vector and study its effect on primary cultures in normal conditions and under hypoxia in vitro.

Materials and Methods. To produce the virus construct, AAV-Syn-EGFP, pDP5, DJ vector, and pHelper plasmids were used. The developed vector was tested on primary hippocampal cells obtained from C57BL/6 mouse embryos on the embryonic day 18 (E18). Infection of primary cultures with the developed virus construct was carried out on the 7^th day of culture development in vitro (7 DIV). The cell viability and spontaneous bioelectrical activity of the cultured cells were assessed on days 1, 3, and 7 after infection. On the 7^th day after infection (14 DIV), spontaneous calcium activity of dissociated cultures was analyzed using functional calcium imaging. To evaluate the neuroprotective properties of the developed construct, in vitro hypoxia modeling was performed on the 14^th day (14 DIV) of cultivation.

Results. The AAV-Syn-BDNF-EGFP adeno-associated virus vector carrying the sequence of the BDNF gene has been developed to increase this neurotrophin expression. Primary hippocampal cells infected by the vector were found to produce increased amounts of BDNF. The developed virus vector did not adversely affect the viability and functional activity of the neural networks in the primary hippocampal cultures. The BDNF overexpression enhanced the neuroprotective potential of cells subjected to oxygen deficiency.

Conclusion. The developed virus containing the BDNF gene sequence increases the endogenous BDNF expression by brain neurons, and thus reduces the death rate of nerve cells in the hypoxia model in vitro.

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Mitroshina E.V., Epifanova E.A., Mishchenko T.A., Yarkov R.S., Babaev A.A., Vedunova M.V. Application of the AAV-Syn-BDNF-EGFP Virus Vector as a Neuroprotective Agent in Modeling Hypoxia in vitro. Sovremennye tehnologii v medicine 2018; 10(2): 47, https://doi.org/10.17691/stm2018.10.2.05