Skip to main content
Log in

Different doses of bone morphogenetic protein 4 promote the expression of early germ cell-specific gene in bone marrow mesenchymal stem cells

  • Report
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

Bone morphogenetic protein (BMP)-4 has a crucial role on primordial germ cells (PGCs) development in vivo which can promote stem cell differentiation to PG-like cells. In this study, we investigated the expression of Mvh as one of the specific genes in primordial germ cells after treatment with different doses of BMP4 on bone mesenchymal stem cells (BMSCs)-derived PGCs. Following isolation of BMSCs from male mouse femur and tibia, cells were cultured in medium for 72 h. Passage 4 murine BMSCs were characterized by CD90, CD105, CD34, and CD45 markers and osteo-adipogenic differentiation. Different doses of BMP4 (0, 0.01, 0.1, 1, 5, 25, 50, and 100 ng/ml) were added to BMSCs for PGCs differentiation during 4-days culture. Viability percent, proliferation rates, and expression of Mvh gene were analyzed by RT-qPCR. Data analysis was done with ANOVA test. CD90+, CD105+, CD34, and CD45 BMSCs were able to differentiate to osteo-adipogenic lineages. The results revealed that proliferation rate and viability percent were raised significantly (p ≤ 0.05) by adding 1, 5, 25 ng/ml of BMP4 and there were decreased to the lowest rate after adding 100 ng/ml BMP4 (p ≤ 0.05). There were significant up regulation (p ≤ 0.05) in Mvh expression between 25, 50, and 100 ng/ml BMP4 with other doses. So the selective dose of BMP-4 for treatment during 4-day culture was 25 ng/ml. The results suggest that addition of 25 ng/ml BMP4 had the best effects based on gene-specific marker expression.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Figure 1.
Figure 2.
Figure 3.
Figure 4.

Similar content being viewed by others

References

  • Aflatoonian B.; Ruban L.; Jones M.; Aflatoonian R.; Fazeli A.; Moore H. D. In vitro post-meiotic germ cell development from human embryonic stem cells. Hum Reprod 24(12): 3150–3159; 2009.

    Article  PubMed  CAS  Google Scholar 

  • Bhatia M.; Bonnet D.; Wu D.; Murdoch B.; Wrana J.; Gallacher L. Bone morphogenetic proteins regulate the developmental program of human hematopoietic stem cells. J Exp Med 189: 1139–1148; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Boroujeni M. B.; Salehnia M.; Valojerdi M. R.; Mowla S. J.; Forouzandeh M.; Hajizadeh E. Comparison of gene expression profiles in erythroid-like cells derived from mouse embryonic stem cells differentiated in simple and co-culture systems. Am J Hematol 83: 109–115; 2008.

    Article  PubMed  CAS  Google Scholar 

  • Buckley S.; Shi W.; Driscoll B.; Ferrario A.; Anderson K.; Warburton D. BMP4 signaling induces senescence and modulates the oncogenic phenotype of A549 lung adenocarcinoma cells. Am J Physiol Lung Cell Mol Physiol 286(1): 81–86; 2004.

    Article  Google Scholar 

  • de Sousa L. S. M.; Roelen B. A.; Monteiro R. M.; Emmens R.; Lin H. Y.; Li E.; Lawson K. A.; Mummery C. L. BMP signaling mediated by ALK2 in the visceral endoderm is necessary for the generation of primordial germ cells in the mouse embryo. Genes Dev 18(15): 1838–1849; 2004.

    Article  Google Scholar 

  • Dudley B. M.; Runyan C.; Takeuchi Y.; Schaible K.; Molyneaux K. BMP signaling regulates PGC numbers and motility in organ culture. Mech Dev 124: 68–77; 2007.

    Article  PubMed  CAS  Google Scholar 

  • Dunn N. R.; Winnier G. E.; Hargett L. K.; Schrick J. J.; Fogo A. B.; Hogan B. L. Haploinsufficient phenotypes in Bmp4 heterozygous null mice and modification by mutations in Gli3 and Alx4. Dev Biol 188(2): 235–247; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Eslaminejad M. B.; Nikmahzar A.; Taghiyar L.; Nadri S.; Massumi M. Murine mesenchymal stem cells isolated by low density primary culture system. Dev Growth Differ 48: 361–370; 2006.

    Article  PubMed  CAS  Google Scholar 

  • Fujiwara Y.; Komiya T.; Kawabata H.; Sato M.; Fujimoto H.; Furusawa M.; Noce T. Isolation of a DEAD-family protein gene that encodes a murine homolog of Drosophila vasa and its specific expression in germ cell lineage. Proc Natl Acad Sci U.S. 91: 12258–12262; 1994.

    Article  CAS  Google Scholar 

  • Fukuda N.; Saitoh M.; Kobayashi N.; Miyazono K. Execution of BMP-4-induced apoptosis by p53-dependent ER dysfunction in myeloma and B-cell hybridoma cells. Oncogene 25: 3509–3517; 2006.

    Article  PubMed  CAS  Google Scholar 

  • Grassinger J.; Simon M.; Mueller G.; Drewel D.; Andreesen R.; Hennemann B. Bone morphogenetic protein (BMP)-7 but not BMP-2 and BMP-4 improves maintenance of primitive peripheral blood-derived hematopoietic progenitor cells (HPC) cultured in serum-free medium supplemented with early acting cytokines. Cytokine 40(3): 165–171; 2007.

    Article  PubMed  CAS  Google Scholar 

  • Hayashi K.; Kobayashi T.; Umino T.; Goitsuka R.; Matsui Y.; Kitamura D. SMAD1 signaling is critical for initial commitment of germ cell lineage from mouse epiblast. Mech Dev 118: 99–109; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Jamous M.; Al-Zoubi A.; Khabaz M. N.; Khaledi R.; Al K. M.; Al-Zoubi Z. Purification of mouse bone marrow-derived stem cells promotes ex vivo neuronal differentiation. Cell Transplant 19(2): 193–202; 2010.

    Article  PubMed  Google Scholar 

  • Kingsley D. M.; Bland A. E.; Grubber J. M.; Marker P. C.; Russell L. B.; Copeland N. G. The mouse short ear skeletal morphogenesis locus is associated with defects in a bone morphogenetic member of the TGF beta superfamily. Cell 71: 399–410; 1992.

    Article  PubMed  CAS  Google Scholar 

  • Kiyono M.; Shibuya M. Bone morphogenetic protein 4 mediates apoptosis of capillary endothelial cells during rat papillary membrane regression. Mol Cell Biol 23(13): 4627–4636; 2003.

    Article  PubMed  CAS  Google Scholar 

  • Kozawa O.; Hatakeyama D.; Uematsu T. Divergent regulation by p44/p42 MAP kinase and p38 MAP kinase of bone morphogenetic protein-4-stimulated osteocalcin synthesis in osteoblasts. J Cell Biochem 84(3): 583–589; 2002.

    Article  PubMed  Google Scholar 

  • Lawson K. A.; Dunn N. R.; Roelen B. A.; Zeinstra L. M.; Davis A. M.; Wright C. V.; Korving J. P.; Hogan B. L. Bmp4 is required for the generation of primordial germ cells in the mouse embryo. Genes Dev 13: 424–436; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Li L.; Xie T. Stem cell niche: Structure and function. Annu Rev Cell Dev Biol 21: 605–631; 2005.

    Article  PubMed  CAS  Google Scholar 

  • Liu S. Y.; Zhang Z. Y.; Song Y. C.; Qiu K. J.; Zhang K. C.; An N. Regulation of BMP4 on the proliferation and differentiation in SVZa neural stem cells. Chin Sci Bull 49(11): 1126–1136; 2004.

    Article  CAS  Google Scholar 

  • Makoolati Z.; Movahedin M.; Forouzandeh-Moghadam M. Effects of different doses of bone morphogenetic protein 4 on viability and proliferation rates of mouse embryonic stem cells. Yakhteh Med J 11: 29–34; 2009.

    Google Scholar 

  • Nagano M. C. In vitro gamete derivation from pluripotent stem cells: Progress and perspective. Biol Reprod 76: 546–551; 2007.

    Article  PubMed  CAS  Google Scholar 

  • Toyooka Y.; Tsunekawa N.; Akasu R.; Noce T. Embryonic stem cells can form germ cells in vitro. Proc Natl Acad Sci USA 100: 11457–11462; 2003.

    Article  PubMed  CAS  Google Scholar 

  • Toyooka Y.; Tsunekawa N.; Takahashi Y.; Matsui Y.; Satoh M.; Noce T. Expression and intracellular localization of mouse VASA-homologue protein during germ cell development. Mech Dev 93: 139–149; 2000.

    Article  PubMed  CAS  Google Scholar 

  • West F. D.; Roche-Rios M. I.; Abraham S.; Rao R. R.; Natrajan M. S.; Bacanamwo M.; Stice S. L. KIT ligand and bone morphogenetic protein signaling enhances human embryonic stem cell to germ-like cell differentiation. Hum Reprod 25(1): 168–178; 2010.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We gratefully thank Dr Makoolati, Mr. Moslem, and Mr Pourbeiranvand for their critical comments and technical support. This study were supported by the Tarbiat Modares University and the Center for International Scientific Cooperation.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Mansoureh Movahedin.

Additional information

Editor: T. Okamoto

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mazaheri, Z., Movahedin, M., Rahbarizadeh, F. et al. Different doses of bone morphogenetic protein 4 promote the expression of early germ cell-specific gene in bone marrow mesenchymal stem cells. In Vitro Cell.Dev.Biol.-Animal 47, 521–525 (2011). https://doi.org/10.1007/s11626-011-9429-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-011-9429-0

Keywords

Navigation