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

The present study aimed to investigate the effects of lentiviral infection of human umbilical cord mesenchymal 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 multipl...

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Main Authors:	CHANG, JING, TANG, LI, LEI, HAN, ZHANG, XIAO-GANG, ZUO, ZHONG, HUANG, WEI, FU, HANG
Format:	Online
Language:	English
Published:	D.A. Spandidos 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4199401/

id	pubmed-4199401
recordtype	oai_dc
spelling	pubmed-41994012014-10-16 Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4 CHANG, JING TANG, LI LEI, HAN ZHANG, XIAO-GANG ZUO, ZHONG HUANG, WEI FU, HANG Articles The present study aimed to investigate the effects of lentiviral infection of human umbilical cord mesenchymal 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. D.A. Spandidos 2014-11 2014-08-21 /pmc/articles/PMC4199401/ /pubmed/25174942 http://dx.doi.org/10.3892/mmr.2014.2505 Text en Copyright © 2014, Spandidos Publications http://creativecommons.org/licenses/by/3.0 This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	CHANG, JING TANG, LI LEI, HAN ZHANG, XIAO-GANG ZUO, ZHONG HUANG, WEI FU, HANG
spellingShingle	CHANG, JING TANG, LI LEI, HAN ZHANG, XIAO-GANG ZUO, ZHONG HUANG, WEI FU, HANG Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4
author_facet	CHANG, JING TANG, LI LEI, HAN ZHANG, XIAO-GANG ZUO, ZHONG HUANG, WEI FU, HANG
author_sort	CHANG, JING
title	Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4
title_short	Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4
title_full	Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4
title_fullStr	Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4
title_full_unstemmed	Effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4
title_sort	effects of lentiviral infection of mesenchymal stem cells on the expression of octamer transcription factor 4
description	The present study aimed to investigate the effects of lentiviral infection of human umbilical cord mesenchymal 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.
publisher	D.A. Spandidos
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4199401/
_version_	1613145559693000704

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

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