Expression of pluripotency transcription factors in human third molar tooth germ derived multipotent mesenchymal stromal cells transfected by plasmid pBud-Sox2-Oct4


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Abstract

In this study, the double expression cassette plasmid, based on pBudCE4 1 vector encoding transcription factors S0X2 and 0CT4 was constructed using standard gene engineering techniques. Expression of recombinant genes was confirmed by immunoblotting. It is shown that genetic modification of multipotent mesenchymal stromal cells (MMSC), isolated from human third molar tooth germs, with resulting recombinant plasmid increases the level of expression both, transcription factors S0X2 and OCT4 in the treated cells, and also transcription factor NAN0G. Analysis of histological sections of subcutaneous Matrigel implants, containing fluorescently labeled MMSC, showed that genetic modification had no effect on cell viability

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About the authors

V. V Solovyeva

Kazan (Volga Region) Federal University

N. L Blatt

Kazan (Volga Region) Federal University

D. S Guseva

Kazan State Medical University; Hannover Medical School

M. E Yalvac

Yeditepe University; Children’s Hospital Columbus

F. Sahin

Yeditepe University

R. R Islamov

Kazan State Medical University

A. A Rizvanov

Kazan (Volga Region) Federal University

References

  1. Takahashi K., Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006;126(4): 663-76.
  2. Maherali N., Sridharan R., Xie W. et al., Directly reprogrammed fibroblasts show global epigenetic remodeling and widespread tissue contribution. Cell Stem Cell 2007; 1(1): 55-70.
  3. Okita K., Ichisaka T., Yamanaka S. Generation of germline-compe-tent induced pluripotent stem cells. Nature 2007; 448(7151): 313-7.
  4. Wernig M., Meissner A., Foreman R. et al. In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature 2007; 448(7151): 318-24.
  5. Nichols J., Zevnik B., Anastassiadis K. et al. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 1998; 95(3): 379-91.
  6. Niwa H., Miyazaki J., Smith A.G. Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat. Gen. 2000; 24(4): 372-6.
  7. Yalvac M.E., Ramazanoglu M., Gumru O.Z. et al. Comparison and optimisation of transfection of human dental follicle cells, a novel source of stem cells, with different chemical methods and electroporation. Neurochem. Res. 2009; 34(7): 1272-7.
  8. Yalvac M.E., Ramazanoglu M., Rizvanov A.A. et al. Isolation and characterization of stem cells derived from human third molar tooth germs of young adults: implications in neo-vascularization, osteo-, adipo- and neurogenesis. Pharmacogenom. J. 2010. 10(2): 105-13.
  9. Kaukua N., Shahidi M.K., Konstantinidou C. et al. Glial origin of mesenchymal stem cells in a tooth model system. Nature 2014; 513(7519): 551-4.
  10. Thesleff I., Sharpe P. Signalling networks regulating dental development. Mech. Dev. 1997; 67(2): 111-23.
  11. Yalvac M.E., Rizvanov A.A., Kilic E. et al. Potential role of dental stem cells in the cellular therapy of cerebral ischemia. Curr. Pharm. Des. 2009; 15(33): 3908-16.
  12. Соловьева В.В., Блатт Н.Л., Шафигуллина А.К. и др. Исследование эндогенной секреции сосудистого эндотелиального фактора роста мультипотентными мезенхимными стромальными клетками из зачатков третьих моляров человека Клеточная трансплантология и тканевая инженерия 2012; Vii(3): 155-58.
  13. Lagarkova M.A., Volchkov P.Y., Lyakisheva A.V. et al. Diverse epigenetic profile of novel human embryonic stem cell lines. Cell Cycle 2006; 5(4): 416-20.
  14. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227(5259): 680-5
  15. Hughes, C.S., Postovit L.M., Lajoie G.A. Matrigel: a complex protein mixture required for optimal growth of cell culture. Proteomics 2010; 10(9): 1886-90.
  16. Rizzino A. Sox2 and 0ct-3/4: a versatile pair of master regulators that orchestrate the self-renewal and pluripotency of embryonic stem cells. Wiley interdiscip. Rev. Syst. Biol. Med. 2009; 1(2): 228-36.
  17. Matin M.M., Walsh J.R., Gokhale P.J. et al. Specific knockdown of 0ct4 and beta2-microglobulin expression by RNA interference in human embryonic stem cells and embryonic carcinoma cells Stem Cells 2004; 22(5): 659-68
  18. Han S.M., Han S.H., Coh Y.R. et al. Enhanced proliferation and differentiation of 0ct4- and Sox2-overexpressing human adipose tissue mesenchymal stem cells. Exp. Mol. Med. 2014; 46: e101.
  19. Guseva, D., Rizvanov A.A., Salafutdinov I.I. et al. 0ver-expression of oct4 and sox2 transcription factors enhances differentiation of human umbilical cord blood cells in vivo Biochem Biophys. Res. Commun. 2014; 451(4): 503-9.
  20. Rodda D.J., Chew J.L., Lim L.H. et al. Transcriptional regulation of nanog by 0CT4 and S0X2 J Biol Chem 2005; 280(26): 24731-7.

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