Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Oncology Letters
      • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Biomedical Reports
      • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • Information for Authors
    • Information for Reviewers
    • Information for Librarians
    • Information for Advertisers
    • Conferences
  • Language Editing
Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • For Authors
    • For Reviewers
    • For Librarians
    • For Advertisers
    • Conferences
  • Language Editing
Login Register Submit
  • This site uses cookies
  • You can change your cookie settings at any time by following the instructions in our Cookie Policy. To find out more, you may read our Privacy Policy.

    I agree
Search articles by DOI, keyword, author or affiliation
Search
Advanced Search
presentation
Biomedical Reports
Join Editorial Board Propose a Special Issue
Print ISSN: 2049-9434 Online ISSN: 2049-9442
Journal Cover
March-2017 Volume 6 Issue 3

Full Size Image

Sign up for eToc alerts
Recommend to Library

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

Medicine International

An International Open Access Journal Devoted to General Medicine.

Journal Cover
March-2017 Volume 6 Issue 3

Full Size Image

Sign up for eToc alerts
Recommend to Library

  • Article
  • Citations
    • Cite This Article
    • Download Citation
    • Create Citation Alert
    • Remove Citation Alert
    • Cited By
  • Similar Articles
    • Related Articles (in Spandidos Publications)
    • Similar Articles (Google Scholar)
    • Similar Articles (PubMed)
  • Download PDF
  • Download XML
  • View XML
Article

Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells

  • Authors:
    • Dae Seong Kim
    • Myoung Woo Lee
    • Tae‑Hee Lee
    • Ki Woong Sung
    • Hong Hoe Koo
    • Keon Hee Yoo
  • View Affiliations / Copyright

    Affiliations: Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea, Department of Laboratory of Cancer and Stem Cell Biology, Plant Engineering Institute, Sejong University, Seoul, Republic of Korea
  • Pages: 300-306
    |
    Published online on: January 19, 2017
       https://doi.org/10.3892/br.2017.845
  • Expand metrics +
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )
Cited By (CrossRef): 0 citations Loading Articles...

This article is mentioned in:



Abstract

The results of clinical trials using mesenchymal stem cells (MSCs) are controversial due to the heterogeneity of human MSCs and differences in culture conditions. In this regard, it is important to identify gene expression patterns according to culture conditions, and to determine how the cells are expanded and when they should be clinically used. In the current study, stemness gene expression was investigated in adipose tissue‑derived MSCs (AT‑MSCs) harvested following culture at different densities. AT‑MSCs were plated at a density of 200 or 5,000 cells/cm2. After 7 days of culture, stemness gene expression was examined by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. The proliferation rate of AT‑MSCs harvested at a low density (~50% confluent) was higher than that of AT‑MSCs harvested at a high density (~90% confluent). Although there were differences in the expression levels of stemness gene, such as octamer‑binding transcription factor 4, nanog homeobox (Nanog), SRY‑box 2, Kruppel like factor 4, v‑myc avian myelocytomatosis viral oncogene homolog (c‑Myc), and lin‑28 homolog A, in the AT‑MSCs obtained from different donors, RT‑qPCR analysis demonstrated differential gene expression patterns according to the cell culture density. Expression levels of stemness genes, particularly Nanog and c‑Myc, were upregulated in AT‑MSCs harvested at a low density (~50% confluent) in comparison to AT‑MSCs from the same donor harvested at a high density (~90% confluent). These results imply that culture conditions, such as the cell density at harvesting, modulate the stemness gene expression and proliferation of MSCs.
View Figures

Figure 1

Figure 2

Figure 3

View References

1 

Tian Y, Deng YB, Huang YJ and Wang Y: Bone marrow-derived mesenchymal stem cells decrease acute graft-versus-host disease after allogeneic hematopoietic stem cells transplantation. Immunol Invest. 37:29–42. 2008. View Article : Google Scholar : PubMed/NCBI

2 

Keyser KA, Beagles KE and Kiem HP: Comparison of mesenchymal stem cells from different tissues to suppress T-cell activation. Cell Transplant. 16:555–562. 2007. View Article : Google Scholar : PubMed/NCBI

3 

Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S and Marshak DR: Multilineage potential of adult human mesenchymal stem cells. Science. 284:143–147. 1999. View Article : Google Scholar : PubMed/NCBI

4 

Sasaki M, Abe R, Fujita Y, Ando S, Inokuma D and Shimizu H: Mesenchymal stem cells are recruited into wounded skin and contribute to wound repair by transdifferentiation into multiple skin cell type. J Immunol. 180:2581–2587. 2008. View Article : Google Scholar : PubMed/NCBI

5 

Colter DC, Class R, DiGirolamo CM and Prockop DJ: Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci USA. 97:3213–3218. 2000. View Article : Google Scholar : PubMed/NCBI

6 

Sekiya I, Larson BL, Smith JR, Pochampally R, Cui JG and Prockop DJ: Expansion of human adult stem cells from bone marrow stroma: Conditions that maximize the yields of early progenitors and evaluate their quality. Stem Cells. 20:530–541. 2002. View Article : Google Scholar : PubMed/NCBI

7 

Lee RH, Hsu SC, Munoz J, Jung JS, Lee NR, Pochampally R and Prockop DJ: A subset of human rapidly self-renewing marrow stromal cells preferentially engraft in mice. Blood. 107:2153–2161. 2006. View Article : Google Scholar : PubMed/NCBI

8 

Estrada JC, Torres Y, Benguría A, Dopazo A, Roche E, Carrera-Quintanar L, Pérez RA, Enríquez JA, Torres R, Ramírez JC, et al: Human mesenchymal stem cell-replicative senescence and oxidative stress are closely linked to aneuploidy. Cell Death Dis. 4:e6912013. View Article : Google Scholar : PubMed/NCBI

9 

Wagner W, Horn P, Castoldi M, Diehlmann A, Bork S, Saffrich R, Benes V, Blake J, Pfister S, Eckstein V, et al: Replicative senescence of mesenchymal stem cells: A continuous and organized process. PLoS One. 3:e22132008. View Article : Google Scholar : PubMed/NCBI

10 

Schellenberg A, Lin Q, Schüler H, Koch CM, Joussen S, Denecke B, Walenda G, Pallua N, Suschek CV, Zenke M, et al: Replicative senescence of mesenchymal stem cells causes DNA-methylation changes which correlate with repressive histone marks. Aging (Albany NY). 3:873–888. 2011. View Article : Google Scholar : PubMed/NCBI

11 

Lee RH, Seo MJ, Pulin AA, Gregory CA, Ylostalo J and Prockop DJ: The CD34-like protein PODXL and alpha6-integrin (CD49f) identify early progenitor MSCs with increased clonogenicity and migration to infarcted heart in mice. Blood. 113:816–826. 2009. View Article : Google Scholar : PubMed/NCBI

12 

Kim DS, Lee MW, Yoo KH, Lee TH, Kim HJ, Jang IK, Chun YH, Kim HJ, Park SJ, Lee SH, et al: Gene expression profiles of human adipose tissue-derived mesenchymal stem cells are modified by cell culture density. PLoS One. 9:e833632014. View Article : Google Scholar : PubMed/NCBI

13 

Lee MW, Kim DS, Yoo KH, Kim HR, Jang IK, Lee JH, Kim SY, Son MH, Lee SH, Jung HL, et al: Human bone marrow-derived mesenchymal stem cell gene expression patterns vary with culture conditions. Blood Res. 48:107–114. 2013. View Article : Google Scholar : PubMed/NCBI

14 

Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP and Hedrick MH: Multilineage cells from human adipose tissue: Implications for cell-based therapies. Tissue Eng. 7:211–228. 2001. View Article : Google Scholar : PubMed/NCBI

15 

Jurgens WJ, Oedayrajsingh-Varma MJ, Helder MN, Zandiehdoulabi B, Schouten TE, Kuik DJ, Ritt MJ and van Milligen FJ: Effect of tissue-harvesting site on yield of stem cells derived from adipose tissue: Implications for cell-based therapies. Cell Tissue Res. 332:415–426. 2008. View Article : Google Scholar : PubMed/NCBI

16 

Lee TH, Song SH, Kim KL, Yi JY, Shin GH, Kim JY, Kim J, Han YM, Lee SH, Lee SH, et al: Functional recapitulation of smooth muscle cells via induced pluripotent stem cells from human aortic smooth muscle cells. Circ Res. 106:120–128. 2010. View Article : Google Scholar : PubMed/NCBI

17 

Martinez-Fernandez A, Nelson TJ, Yamada S, Reyes S, Alekseev AE, Perez-Terzic C, Ikeda Y and Terzic A: iPS programmed without c-MYC yield proficient cardiogenesis for functional heart chimerism. Circ Res. 105:648–656. 2009. View Article : Google Scholar : PubMed/NCBI

18 

Mauritz C, Schwanke K, Reppel M, Neef S, Katsirntaki K, Maier LS, Nguemo F, Menke S, Haustein M, Hescheler J, et al: Generation of functional murine cardiac myocytes from induced pluripotent stem cells. Circulation. 118:507–517. 2008. View Article : Google Scholar : PubMed/NCBI

19 

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

20 

Zhang F, Citra F and Wang DA: Prospects of induced pluripotent stem cell technology in regenerative medicine. Tissue Eng Part B Rev. 17:115–124. 2011. View Article : Google Scholar : PubMed/NCBI

21 

Liu YH, Karra R and Wu SM: Cardiovascular stem cells in regenerative medicine: Ready for prime time? Drug Discov Today Ther Strateg. 5:201–207. 2008. View Article : Google Scholar : PubMed/NCBI

22 

Uccelli A, Moretta L and Pistoia V: Mesenchymal stem cells in health and disease. Nat Rev Immunol. 8:726–736. 2008. View Article : Google Scholar : PubMed/NCBI

23 

Murphy MB, Moncivais K and Caplan AI: Mesenchymal stem cells: Environmentally responsive therapeutics for regenerative medicine. Exp Mol Med. 45:e542013. View Article : Google Scholar : PubMed/NCBI

24 

Melief SM, Zwaginga JJ, Fibbe WE and Roelofs H: Adipose tissue-derived multipotent stromal cells have a higher immunomodulatory capacity than their bone marrow-derived counterparts. Stem Cells Transl Med. 2:455–463. 2013. View Article : Google Scholar : PubMed/NCBI

25 

Smith JR, Pochampally R, Perry A, Hsu SC and Prockop DJ: Isolation of a highly clonogenic and multipotential subfraction of adult stem cells from bone marrow stroma. Stem Cells. 22:823–831. 2004. View Article : Google Scholar : PubMed/NCBI

26 

Pricola KL, Kuhn NZ, Haleem-Smith H, Song Y and Tuan RS: Interleukin-6 maintains bone marrow-derived mesenchymal stem cell stemness by an ERK1/2-dependent mechanism. J Cell Biochem. 108:577–588. 2009. View Article : Google Scholar : PubMed/NCBI

27 

Han SM, Han SH, Coh YR, Jang G, Ra J Chan, Kang SK, Lee HW and Youn HY: Enhanced proliferation and differentiation of Oct4- and Sox2-overexpressing human adipose tissue mesenchymal stem cells. Exp Mol Med. 46:e1012014. View Article : Google Scholar : PubMed/NCBI

28 

Go MJ, Takenaka C and Ohgushi H: Forced expression of Sox2 or Nanog in human bone marrow derived mesenchymal stem cells maintains their expansion and differentiation capabilities. Exp Cell Res. 314:1147–1154. 2008. View Article : Google Scholar : PubMed/NCBI

29 

Alison MR and Islam S: Attributes of adult stem cells. J Pathol. 217:144–160. 2009. View Article : Google Scholar : PubMed/NCBI

30 

Iovino N and Cavalli G: Rolling ES cells down the Waddington landscape with Oct4 and Sox2. Cell. 145:815–817. 2011. View Article : Google Scholar : PubMed/NCBI

31 

Liu TM, Wu YN, Guo XM, Hui JH, Lee EH and Lim B: Effects of ectopic Nanog and Oct4 overexpression on mesenchymal stem cells. Stem Cells Dev. 18:1013–1022. 2009. View Article : Google Scholar : PubMed/NCBI

32 

Tondreau T, Meuleman N, Delforge A, Dejeneffe M, Leroy R, Massy M, Mortier C, Bron D and Lagneaux L: Mesenchymal stem cells derived from CD133-positive cells in mobilized peripheral blood and cord blood: Proliferation, Oct4 expression, and plasticity. Stem Cells. 23:1105–1112. 2005. View Article : Google Scholar : PubMed/NCBI

33 

Greco SJ, Liu K and Rameshwar P: Functional similarities among genes regulated by OCT4 in human mesenchymal and embryonic stem cells. Stem Cells. 25:3143–3154. 2007. View Article : Google Scholar : PubMed/NCBI

34 

Tsai CC, Su PF, Huang YF, Yew TL and Hung SC: Oct4 and Nanog directly regulate Dnmt1 to maintain self-renewal and undifferentiated state in mesenchymal stem cells. Mol Cell. 47:169–182. 2012. View Article : Google Scholar : PubMed/NCBI

35 

Evans PM, Zhang W, Chen X, Yang J, Bhakat KK and Liu C: Kruppel-like factor 4 is acetylated by p300 and regulates gene transcription via modulation of histone acetylation. J Biol Chem. 282:33994–34002. 2007. View Article : Google Scholar : PubMed/NCBI

36 

Scheper W and Copray S: The molecular mechanism of induced pluripotency: A two-stage switch. Stem Cell Rev. 5:204–223. 2009. View Article : Google Scholar : PubMed/NCBI

37 

Varlakhanova NV, Cotterman RF, deVries WN, Morgan J, Donahue LR, Murray S, Knowles BB and Knoepfler PS: myc maintains embryonic stem cell pluripotency and self-renewal. Differentiation. 80:9–19. 2010. View Article : Google Scholar : PubMed/NCBI

38 

Kern S, Eichler H, Stoeve J, Klüter H and Bieback K: Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 24:1294–1301. 2006. View Article : Google Scholar : PubMed/NCBI

Related Articles

  • Abstract
  • View
  • Download
  • Twitter
Copy and paste a formatted citation
Spandidos Publications style
Kim D, Lee M, Lee TH, Sung K, Koo H and Yoo K: Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells. Biomed Rep 6: 300-306, 2017.
APA
Kim, D., Lee, M., Lee, T., Sung, K., Koo, H., & Yoo, K. (2017). Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells. Biomedical Reports, 6, 300-306. https://doi.org/10.3892/br.2017.845
MLA
Kim, D., Lee, M., Lee, T., Sung, K., Koo, H., Yoo, K."Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells". Biomedical Reports 6.3 (2017): 300-306.
Chicago
Kim, D., Lee, M., Lee, T., Sung, K., Koo, H., Yoo, K."Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells". Biomedical Reports 6, no. 3 (2017): 300-306. https://doi.org/10.3892/br.2017.845
Copy and paste a formatted citation
x
Spandidos Publications style
Kim D, Lee M, Lee TH, Sung K, Koo H and Yoo K: Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells. Biomed Rep 6: 300-306, 2017.
APA
Kim, D., Lee, M., Lee, T., Sung, K., Koo, H., & Yoo, K. (2017). Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells. Biomedical Reports, 6, 300-306. https://doi.org/10.3892/br.2017.845
MLA
Kim, D., Lee, M., Lee, T., Sung, K., Koo, H., Yoo, K."Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells". Biomedical Reports 6.3 (2017): 300-306.
Chicago
Kim, D., Lee, M., Lee, T., Sung, K., Koo, H., Yoo, K."Cell culture density affects the stemness gene expression of adipose tissue‑derived mesenchymal stem cells". Biomedical Reports 6, no. 3 (2017): 300-306. https://doi.org/10.3892/br.2017.845
Follow us
  • Twitter
  • LinkedIn
  • Facebook
About
  • Spandidos Publications
  • Careers
  • Cookie Policy
  • Privacy Policy
How can we help?
  • Help
  • Live Chat
  • Contact
  • Email to our Support Team