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

doi:10.3892/mmr.2014.2505

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

Authors:
- Jing Chang
- Li Tang
- Han Lei
- Xiao-Gang Zhang
- Zhong Zuo
- Wei Huang
- Hang Fu
View Affiliations

Affiliations: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: August 21, 2014 https://doi.org/10.3892/mmr.2014.2505
Pages: 2249-2254
Copyright: © Chang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

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.

View Figures

View References

1	Takahashi K and Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 126:663–676. 2006. View Article : Google Scholar : PubMed/NCBI
2	Okita K, Ichisaka T and Yamanaka S: Generation of germline-competent induced pluripotent stem cells. Nature. 448:313–317. 2007. View Article : Google Scholar : PubMed/NCBI
3	Lin SL, Chang DC, Lin CH, Ying SY, Leu D and Wu DT: Regulation of somatic cell reprogramming through inducible mir-302 expression. Nucleic Acids Res. 39:1054–1065. 2011. View Article : Google Scholar : PubMed/NCBI
4	Card DA, Hebbar PB, Li L, et al: Oct4/Sox2-regulated mir-302 targets cyclin D1 in human embryonic stem cells. Mol Cell Biol. 28:6426–6438. 2008. View Article : Google Scholar : PubMed/NCBI
5	Zovoilis A, Pantazi A, Smoraq L, et al: Embryonic stem cell-related miRNAs are involved in differentiation of pluripotent cells originating from the germ line. Mol Hum Reprod. 16:793–803. 2010. View Article : Google Scholar : PubMed/NCBI
6	Schmidt D, Mol A, Odermatt B, et al: Engineering of biologically active living heart valve leaflets using human umbilical cord-derived progenitor cells. Tissue Eng. 12:3223–3232. 2006. View Article : Google Scholar : PubMed/NCBI
7	Sarugaser R, Lickorish D, Baksh D, Hosseini MM and Davies JE: Human umbilical cord perivascular (HUCPV) cells: A source of mesenchymal progenitors. Stem Cells. 23:220–229. 2005.PubMed/NCBI
8	Boyer LA, Lee TI, Cole MF, et al: Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 122:947–956. 2005. View Article : Google Scholar : PubMed/NCBI
9	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001.
10	Wang H, Yang Y, Ho G, et al: Programming of human umbilical cord mesenchymal stem cells in vitro to promote pancreatic gene expression. Mol Med Rep. 8:769–774. 2013.PubMed/NCBI
11	Phermthai T, Odglun Y, Julavijitphong S, et al: A novel method to derive amniotic fluid stem cells for therapeutic purposes. BMC Cell Biol. 11:792010. View Article : Google Scholar : PubMed/NCBI
12	Troyer DL and Weiss ML: Wharton’s jelly-derived cells are a primitive stromal cell population. StemCells. 26:591–599. 2008.
13	Kim J, Lee Y, Kim H, et al: Human amniotic fluid-derived stem cells have characteristics of multipotent stem cells. Cell Prolif. 40:75–90. 2007.PubMed/NCBI
14	Schneider RK, Püllen A, Kramann R, et al: Long-term survival and characterisation of human umbilical cord-derived mesenchymal stem cells on dermal equivalents. Differentiation. 79:182–193. 2010. View Article : Google Scholar : PubMed/NCBI
15	Atlasi Y, Mowla SJ, Ziaee SA and Bahrami AR: OCT-4, an embryonic stem cell marker, is highly expressed in bladder cancer. Int J Cancer. 120:1598–1602. 2007. View Article : Google Scholar : PubMed/NCBI
16	Hochedlinger K, Yamada Y, Beard C and Jaenisch R: Ectopic expression of Oct-4 blocks progenitor-cell differentiation and causes dysplasia in epithelial tissues. Cell. 121:465–477. 2005. View Article : Google Scholar
17	Mueller T, Luetzkendorf J, Nerger K, Schmoll HJ and Mueller LP: Analysis of OCT4 expression in an extended panel of human tumor cell lines from multiple entities and in human mesenchymal stem cells. Cell Mol Life Sci. 66:495–503. 2009. View Article : Google Scholar : PubMed/NCBI
18	Kaltz N, Funari A, Hippauf S, et al: In vivo osteoprogenitor potency of human stromal cells from different tissues does not correlate with expression of POU5F1 or its pseudogenes. Stem Cells. 26:2419–2424. 2008. View Article : Google Scholar
19	Lengner CJ, Camargo FD, Hochedlinger K, et al: Oct4 expression is not required for mouse somatic stem cell self-renewal. Cell Stem Cell. 1:403–415. 2007. View Article : Google Scholar : PubMed/NCBI
20	Carlin R, Davis D, Weiss M, Schultz B and Troyer D: Expression of early transcription factors Oct-4, Sox-2 and Nanog by porcine umbilical cord (PUC) matrix cells. Reprod Biol Endocrinol. 4:82006. View Article : Google Scholar : PubMed/NCBI
21	Can A and Karahuseyinoglu S: Concise review: human umbilical cord stroma with regard to the source of fetus-derived stem cells. Stem Cells. 25:2886–2895. 2007.
22	Weinberg MS, Barichievy S, Schaffer L, Han J and Morris KV: An RNA targeted to the HIV-1 LTR promoter modulates indiscriminate off-target gene activation. Nucleic Acids Res. 35:7303–7312. 2007. View Article : Google Scholar : PubMed/NCBI
23	Cockrell AS and Kafri T: Gene delivery by lentivirus vectors. Mol Biotechnol. 36:184–204. 2007. View Article : Google Scholar
24	Peng X, Zhang X and Zeng B: Locally administered lentivirus-mediated siRNA inhibits wear debris-induced inflammation in murine air pouch model. Biotechnol Lett. 30:1923–1929. 2008. View Article : Google Scholar : PubMed/NCBI
25	Miyoshi N, Ishii H, Nagai K, et al: Defined factors induce reprogramming of gastrointestinal cancer cells. Proc Natl Acad Sci USA. 107:40–45. 2010. View Article : Google Scholar : PubMed/NCBI