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

Chang J, Tang L, Lei H, Zhang XG, Zuo Z, Huang W, Fu H - Mol Med Rep (2014)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.

ABSTRACT
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.

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Expression of the octamer transcription factor 4 gene (Oct4), as assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). There is no statistically significant difference in the relative expression level of Oct4 between two and eight weeks of culture.
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f6-mmr-10-05-2249: Expression of the octamer transcription factor 4 gene (Oct4), as assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). There is no statistically significant difference in the relative expression level of Oct4 between two and eight weeks of culture.

Mentions: The Oct4 expression level was defined as 1 in the lentivirus-infected group of hUCMSCs after two weeks of culture, and thus, the level of Oct4 was estimated at 0.9075±0.0124 after eight weeks of culture (Fig. 6). The relative expression of Oct4 was not significantly different between cells cultured for two and eight weeks (P>0.05).


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

Chang J, Tang L, Lei H, Zhang XG, Zuo Z, Huang W, Fu H - Mol Med Rep (2014)

Expression of the octamer transcription factor 4 gene (Oct4), as assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). There is no statistically significant difference in the relative expression level of Oct4 between two and eight weeks of culture.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4199401&req=5

f6-mmr-10-05-2249: Expression of the octamer transcription factor 4 gene (Oct4), as assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). There is no statistically significant difference in the relative expression level of Oct4 between two and eight weeks of culture.
Mentions: The Oct4 expression level was defined as 1 in the lentivirus-infected group of hUCMSCs after two weeks of culture, and thus, the level of Oct4 was estimated at 0.9075±0.0124 after eight weeks of culture (Fig. 6). The relative expression of Oct4 was not significantly different between cells cultured for two and eight weeks (P>0.05).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus