Volume 16, Issue 8 (August 2018)                   IJRM 2018, 16(8): 507-518 | Back to browse issues page


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Gholamitabar Tabari M, Jorsaraei S G A, Ghasemzadeh-Hasankolaei M, Ahmadi A A, Mahdinezhad Gorji N. Evaluation of specific germ cell genes expression in mouse embryonic stem cell-derived germ cell like cells treated with bone morphogenetic protein 4 in vitro. IJRM. 2018; 16 (8) :507-518
URL: http://journals.ssu.ac.ir/ijrmnew/article-1-1177-en.html
1- Infertility and Reproductive Health Research Center, Health Research Institute, Babol University of Medical Science, Babol, Iran, Iran
2- Infertility and Reproductive Health Research Center, Health Research Institute, Babol University of Medical Science, Babol, Iran., Iran , alijorsara@yahoo.com
3- Infertility and Reproductive Health Research Center, Health Research Institute, Babol University of Medical Science, Babol, Iran., Iran
Abstract:   (131 Views)
Background: Bone morphogenetic protein 4 (BMP4) is a significant signaling molecule that involves in initiating of differentiation and performs multifunctional effects on embryonic stem cells (ESCs) and embryos.
Objective: The goal of the present study was to evaluate an in vitro differentiation model of mouse embryonic stem cells into germ cells, using BMP4.
Materials and Methods: in this experimental study, we used Oct4-GFP mouse ESCs to form embryoid body (EB) aggregations for two days. Then, single cells from EB were cultured for four days with BMP4. Using MTT assay and gene expression levels for evaluation of Mvh and Riken by real-time RT-PCR of six concentrations, 12.5 ng/ ml BMP4 was determined as an optimized dose. Then, the expression level of Fkbp6, Mov10l1, 4930432K21Rik, Tex13, Mvh, Scp3, Stra8, Oct4 were evaluated. Flow cytometry and immunostaning were used to confirm the findings of the real-time RT-PCR.
Results: In the +BMP4 group, the genes encoding Riken (p≤0.001) and Mvh (p≤0.001) were found to be increased with significant differences compared with the control group. Mov10l1 (p=0.22), Tex13 (p=0.10), Fkbp6 (p=0.90), Scp3 (p=0.61) and Stra8 (p=0.08) were up-regulated without significance differences compared with control group. Flow cytometry analysis showed that the mean number of Mvh-positive cells in the +BMP4 group was greater when compared with ESCs, -BMP4 and EB groups (p=0.03, p≤0.001, p=0.02, respectively).
Conclusion: Down-regulation of Oct4, expression of germ cells genes and meiosis markers expression raise this hypothesis that ESCs were differentiated by BMP4, and may be introduced into the first meiosis as germ cell-like cells.
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Type of Study: Original Article |
Received: 2018/08/27 | Accepted: 2018/08/27 | Published: 2018/08/27

References
1. Ginsburg M, Snow MH, McLaren A. Primordial germ cells in the mouse embryo during gastrulation. Development 1990; 110: 521-528.
2. Daley GQ. Gametes from embryonic stem cells: a cup half empty or half full? Science 2007; 316: 409-410. [DOI:10.1126/science.1138772]
3. Childs AJ, Saunders PT, Anderson RA. Modelling germ cell development in vitro. Mol Hum Reprod 2008; 14: 501-511. [DOI:10.1093/molehr/gan042]
4. Desjardins C, Larry L, Ewing LL. Cell and Molecular Biology of Testis. 1st Ed. Oxford University Press; UK; 1993.
5. Byskov AG, Andersen CY, Nordholm L, Thogersen H, Xia G, Wassmann O, et al. Chemical structure of sterols that activate oocyte meiosis. Nature 1995; 374: 559-562. [DOI:10.1038/374559a0]
6. McLaren A, Southee D. Entry of mouse embryonic germ cells into meiosis. Dev Biol 1997; 187: 107-113. [DOI:10.1006/dbio.1997.8584]
7. Matsui Y, Toksoz D, Nishikawa S, Nishikawa SI, Williams D, Zsebo K et al. Effect of steel factor and leukemia inhibitory factor on murine primordial germ cells in culture. Nature 1991; 353: 750-752. [DOI:10.1038/353750a0]
8. Kawase E, Yamamoto H, Hashimoto K, Nakatsuji N. Tumor necrosis factor-b (TNF-b) stimulates proliferation of mouse primordial germ cells in culture. Dev Biol 1994; 161: 91-95. [DOI:10.1006/dbio.1994.1011]
9. Kocer A, Reichmann J, Best D, Adams IR. Germ cell sex determination in mammals. Mol Hum Reprod 2009; 15: 205-213. [DOI:10.1093/molehr/gap008]
10. Ying Y, Zhao GQ. Cooperation of endoderm-derived BMP2 and extraembryonic ectoderm-derived BMP4 in primordial germ cell generation in the mouse. Dev Biol 2001; 232: 484-492. [DOI:10.1006/dbio.2001.0173]
11. Young JC, Dias VL, Loveland KL. Defining the window of germline genesis in vitro from murine embryonic stem cells. Biol Reprod 2010; 82: 390-401. [DOI:10.1095/biolreprod.109.078493]
12. Tilgner K, Atkinson SP, Golebiewska A, Stojkovic M, Lako M, Armstrong L. Isolation of primordial germ cells from differentiating human embryonic stem cells. Stem Cells 2008; 26: 3075-3085. [DOI:10.1634/stemcells.2008-0289]
13. West FD, Roche-Rios MI, Abraham S, Rao RR, Natrajan MS, Bacanamwo M, et al. KIT ligand and bone morphogenetic protein signaling enhances human embryonic stem cell to germ-like cell differentiation. Hum Reprod 2010; 25: 168-178. [DOI:10.1093/humrep/dep338]
14. Zhang P, Li J, Tan Z, Wang C, Liu T, Chen L, et al. Short-term BMP-4 treatment initiates mesoderm induction in human embryonic stem cells. Blood 2008; 111: 1933-1941. [DOI:10.1182/blood-2007-02-074120]
15. Bowles J, Koopman P. Retinoic acid, meiosis and germ cell fate in mammals. Development 2007; 134: 3401-3411. [DOI:10.1242/dev.001107]
16. Masui S, Nakatake Y, Toyooka Y, Shimosato D, Yagi R, Takahashi K, et al. Pluripotency governed by Sox2 via regulation of Oct3/4 expression in mouse embryonic stem cells. Nat Cell Biol 2007; 9: 625-635. [DOI:10.1038/ncb1589]
17. Wang PJ, McCarrey JR, Yang F, Page DC. An abundance of X-linked gen expressed in spermatogonia. Nat Genet 2001; 27: 422-426. [DOI:10.1038/86927]
18. Kwon JT, Jin S, Choi H, Kim J, Jeong J, Kim J, et al. Identification and characterization of germ cell genes expressed in the F9 testicular teratoma stem cell line. PLoS One 2014; 9: e103837. [DOI:10.1371/journal.pone.0103837]
19. Leitch HG, Tang WW, Surani MA. Primordial germ-cell development and epigenetic reprogramming inmammals. Curr Top Dev Biol 2013; 104: 149-187. [DOI:10.1016/B978-0-12-416027-9.00005-X]
20. Sabour D, Arauzo-Bravo MJ, Hubner K, Greber B, Gentile L, Stehling M, et al. Identification of genes specific to mouse primordial germ cells through dynamic global gene expression. Hum Mol Genet 2011; 20: 115-125. [DOI:10.1093/hmg/ddq450]
21. Mazaheri Z, Movahedin M, Rahbarizadeh F, Amanpour S. 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 Anim 2011; 47: 521-525. [DOI:10.1007/s11626-011-9429-0]
22. Aflatoonian B, Ruban L, Jones M, Aflatoonian R, Fazeli A, Moore HD. In vitro post-meiotic germ cell development from human embryonic stem cells. Hum Reprod 2009; 24: 3150-3159. [DOI:10.1093/humrep/dep334]
23. Pesce M, Klinger GF, De Felici M. Derivation in culture of primordial germ cells from cells of the mouse epiblast: phenotypic induction and growth control by Bmp4 signalling. Mech Dev 2002; 112: 15-24. [DOI:10.1016/S0925-4773(01)00624-4]
24. Ying Y, Qi X, Zhao GQ. Induction of primordial germ cells from murine epiblasts by synergistic action of BMP4 and BMP8B signaling pathways. Proc Natl Acad Sci USA 2001; 98: 7858-7862. [DOI:10.1073/pnas.151242798]
25. Hernandez-Garcia D, Castro-Obregon S, Gomez-Lopez S, Valencia C, Covarrubias L. Cell death activation during cavitation of embryoid bodies is mediated by hydrogen peroxide. Exp Cell Res 2008; 314: 2090-2099. [DOI:10.1016/j.yexcr.2008.03.005]
26. 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 2000; 93: 139-149. [DOI:10.1016/S0925-4773(00)00283-5]
27. Wang PJ, Page DC, McCarrey JR. Differential expression of sex-linked and autosomal germ-cell-specific genes during spermatogenesis in the mouse. Hum Mol Genet 2005; 14: 2911-2918. [DOI:10.1093/hmg/ddi322]
28. Ikadai H, Noguchi J, Yoshida M, Imamichi T. An aspermia rat mutant (as/as) with spermatogenic failure at meiosis. J Vet Med Sci 1992; 54: 745-749. [DOI:10.1292/jvms.54.745]
29. Tanaka SS, Toyooka Y, Akasu R, Katoh-Fukui Y, Nakahara Y, Suzuki R, et al.The mouse homolog of Drosophila Vasa is required for the development of male germ cells. Genes Dev 2000; 14: 841-853.
30. Wang PJ, Page DC, McCarrey JR. Differential expression of sex-linked and autosomal germ-cell- specific genes during spermatogenesis in the mouse. Hum Mol Genet 2005; 14: 2911-2918. [DOI:10.1093/hmg/ddi322]
31. Anderson RA, Fulton N, Cowan G, Coutts S, Saunders PT. Conserved and divergent patterns of expression of DAZL, VASA and OCT4 in the germ cells of the human fetal ovary and testis. BMC Dev Biol 2007; 7: 136. [DOI:10.1186/1471-213X-7-136]
32. Pesce M, Wang X, Wolgemuth DJ, Scholer H. Differential expression of the Oct-4 transcription factor during mouse germ cell differentiation. Mech Dev 1998; 71: 89-98. [DOI:10.1016/S0925-4773(98)00002-1]
33. Yuan L, Liu JG, Zhao J, Brundell E, Daneholt B, Hoog C. The murine SCP3 gene is required for synaptonemal complex assembly, chromosome synapsis, and male fertility. Mol Cell 2000; 5: 73-83. [DOI:10.1016/S1097-2765(00)80404-9]
34. Anderson EL, Baltus AE, Roepers-Gajadien HL, Hassold TJ, de Rooij DG, van Pelt AM, et al. Stra8 and its inducer, retinoic acid, regulate meiotic initiation in both spermatogenesis and oogenesis in mice. Proc Natl Acad Sci USA 2008; 105: 14976-14980. [DOI:10.1073/pnas.0807297105]

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