Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4

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Abstract

The present study aimed to investigate the effects of lentiviral infection of human umbilical cord mesen-chymal stem cells (hUCMSCs) on the expression of octamer transcription factor 4 (Oct4). hUCMSCs were infected with lentivirus carrying the green fluorescent protein gene (GFP) at different multiplicities of infection (MOI), and the optimal MOI was determined by flow cytometry; the proliferation of non infected and GFP-carrying lentivirus infected hUCMSCs was evaluated by the MTT assay; and the expression of the Oct4 gene was measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunofluorescence staining in hUCMSCs cultured in vitro for eight weeks. Positive GFP staining of hUCMSCs was estimated at >75% at 48 h following infection with the GFP-carrying lentivirus (MOI = 20); no effect on hUCMSC proliferation was detected by the MTT assay following the infection; immunofluorescence analysis detected positive Oct4 expression in the cell nuclei at two and eight weeks of culture, while the relative expression of Oct4 assessed by qRT-PCR was 0.9075±0.0124. The GFP gene carried by the lentivirus was successfully expressed in hUCMSCs and had no significant effect on Oct4 expression, which lays a solid foundation for future studies investigating gene functions via the use of exogenous markers.

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Chang, J., Tang, L., Lei, H., Zhang, X. G., Zuo, Z., Huang, W., & Fu, H. (2014). Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4. Molecular Medicine Reports, 10(5), 2249–2254. https://doi.org/10.3892/mmr.2014.2505

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