Human Wharton’s jelly-derived mesenchymal stem cells express oocyte developmental genes during co-culture with placental cells

Document Type: Original Article


1 Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran

2 Department of Tissue engineering, School of Advanced Technologies, Tehran University of Medical Sciences, Tehran, Iran

3 Department of Anatomy, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran

4 Department of Anatomy, Faculty of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran


Objective(s): The present day challenge is how to obtain germ cells from stem cells to treat patients with cancer and infertility. Much more efforts have been made to develop a procedure for attaining germ cells in vitro. Recently, human umbilical cord-derived mesenchymal stem cells (HUMSCs) have been introduced with higher efficacy for differentiation. In this work, we tried to explore the efficacy of HUMSCs and some effective products of placental cells such as transforming growth factors. This study is aimed to optimize a co-culture condition for HUMSCs with placental cells to obtain primordial germ cells (PGCs) and reach into oocyte-like cells in vitro.
Materials and Methods: In this experimental study, HUMSCs and placental cells were co-cultured for 14 days without any external inducer in vitro. Then HUMSCs were assessed for expression of PGC markers; Octamer-binding transcription factor 4(OCT4),Tyrosine-protein kinase Kit (CKIT), Stage specific embryonic antigen 4  (SSEA4) , DEAD (Asp-Glu-Ala-Asp) box polypeptide 4( DDX4) and oocyte specific markers; Growth differentiation factor-9( GDF9), Zona pellucida glycoprotein 3(ZP3). The pertinent markers were assessed by immunocytochemistry and Q-PCR.
Results: Co-cultured HUMSCs with placental cells (including amniotic and chorionic cells) presented Oct4 and DDX4, primordial germ cells specific markers significantly, but increment in expression of oocyte-like cell specific markers, GDF9 and ZP3 did not reach to statistically significant threshold.  
Conclusion: Placental cell supplementsTransforming growth factor (TGF α, β) and basic fibroblast growth factor (bFGF) in a co-culture model can provide proper environment for induction of HUMSCs into PGCs and expression of oocyte-like markers.


1. Mascarenhas MN, Flaxman SR, Boerma T, Vanderpoel S, Stevens GA. National, regional, and global trends in infertility prevalence since 1990: a systematic analysis of 277 health surveys. PLoS Med 2012; 9:e1001356.

2. Ziebe S, Loft A, Petersen JH, Andersen AG, Lindenberg S, Petersen K, et al. Embryo quality and developmental potential is compromised by age. Acta Obstet Gynecol Scand 2001; 80:169-174.

3. Marques-Mari AI, Lacham-Kaplan O, Medrano JV, Pellicer A, Simon C. Differentiation of germ cells and gametes from stem cells. Hum Reprod Update 2009; 15:379-390.

4. Hubner K, Fuhrmann G, Christenson LK, Kehler J, Reinbold R, De La Fuente R, et al. Derivation of oocytes from mouse embryonic stem cells. Science 2003; 300:1251-1256.

5. Lacham-Kaplan O, Chy H, Trounson A. Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes. Stem Cells 2006; 24:266-273.

6. Chen J, Lu Z, Cheng D, Peng S, Wang H. Isolation and characterization of porcine amniotic fluid-derived multipotent stem cells. PLoS One 2011; 6:e19964.

7. Nayernia K, Lee JH, Drusenheimer N, Nolte J, Wulf G, Dressel R, et al. Derivation of male germ cells from bone marrow stem cells. Lab Invest 2006; 86:654-663.

8. Dyce PW, Li J. From skin cells to ovarian follicles? Cell Cycle 2006; 5:1371-1375.

9. Johnson J, Bagley J, Skaznik-Wikiel M, Lee HJ, Adams GB, Niikura Y, et al. Oocyte generation in adult mammalian ovaries by putative germ cells in bone marrow and peripheral blood. Cell 2005; 122:303-315.

10. Nikoozad Z, Ghorbanian MT, Rezaei A. Comparison of the liver function and hepatic specific genes expression in cultured mesenchymal stem cells and hepatocytes. Iran J Basic Med Sci 2014; 17:27-33.

11. Salehinejad P, Alitheen NB, Ali AM, Omar AR, Mohit M, Janzamin E, et al. Comparison of different methods for the isolation of mesenchymal stem cells from human umbilical cord Wharton's jelly. In Vitro Cell Dev Biol Anim 2012; 48:75-83.

12. Parolini O, Alviano F, Bagnara GP, Bilic G, Bühring HJ, Evangelista M, et al. Concise review: isolation and characterization of cells from human term placenta: outcome of the first international workshop on placenta derived stem cells. Stem Cells 2008; 26:300-311.

13. Denker H. Embryonic stem cells: An exciting field for basic research and tissue engineering, but also an ethical dilemma? Cells Tissues Organs 1999; 165:246-249.

14. Romano G. Stem cell transplantation therapy: controversy over ethical issues and clinical relevance. Drug News Perspect 2004; 17:637-645.

15. Ma L, Feng XY, Cui BL, Law F, Jiang XW, Yang LY, et al. Human umbilical cord Wharton's Jelly-derived mesenchymal stem cells differentiation into nerve-like cells. Chin Med J (Engl) 2005; 118:1987-1993.

16. Troyer DL, Weiss ML. Wharton's jelly-derived cells are a primitive stromal cell population. Stem Cells 2008; 26:591-599.

17. Wang L, Ott L, Seshareddy K, Weiss ML, Detamore MS. Musculoskeletal tissue engineering with human umbilical cord mesenchymal stromal cells. Regen Med 2011; 6:95-109.

18. Hua J, Qiu P, Zhu H, Cao H, Wang F, Li W. Multipotent mesenchymal stem cells (MSCs) from human umbilical cord: potential differentiation of germ cells. Afr J Biochem Res 2011; 5:113-123.

19. Hass R, Kasper C, Bohm S, Jacobs R. Different populations and sources of human mesenchymal stem cells (MSC): A comparison of adult and neonatal tissue-derived MSC. Cell Commun Signal 2011; 9:12.

20. Huang P, Lin LM, Wu XY, Tang QL, Feng XY, Lin GY, et al. Differentiation of human umbilical cord Wharton's jelly-derived mesenchymal stem cells into germ-like cells in vitro. J Cell Biochem 2010; 109:747-754.

21. Huang S, Feng C, Wu Y, Yang S, Ma K, Wu X, et al. Dissimilar characteristics of umbilical cord mesenchymal stem cells from donors of different ages. Cell Tissue Bank 2013; 14:707-713.

22. Toyooka Y, Tsunekawa N, Akasu R, Noce T. Embryonic stem cells can form germ cells in vitro. Proc Natl Acad Sci U S A 2003; 100:11457-11462.

23. Miki T, Strom SC. Amnion-derived pluripotent/ multipotent stem cells. Stem Cell Rev 2006; 2:133-142.

24. Park Y, Lee SJ, Choi IY, Lee SR, Sung HJ, Kim JH, et al. The efficacy of human placenta as a source of the universal feeder in human and mouse pluripotent stem cell culture. Cell Reprogram 2010; 12:315-328.

25. Miyamoto K, Hayashi K, Suzuki T, Ichihara S, Yamada T, Kano Y, et al. Human placenta feeder layers support undifferentiated growth of primate embryonic stem cells. Stem Cells 2004; 22:433-440.

26. Nilsson E, Parrott JA, Skinner MK. Basic fibroblast growth factor induces primordial follicle develop-ment and initiates folliculogenesis. Mol Cell Endocrinol 2001; 175:123-130.

27. Zhang Y, Li C, Jiang X, Zhang S, Wu Y, Liu B, et al. Human placenta-derived mesenchymal progenitor cells support culture expansion of long-term culture-initiating cells from cord blood CD34< sup>+ cells. Exp Hematol 2004; 32:657-664.

28. Gordon KJ, Blobe GC. Role of transforming growth factor-beta superfamily signaling pathways in human disease. Biochim Biophys Acta 2008; 1782:197-228.

29. Walz TM, Malm C, Nishikawa BK, Wasteson A. Transforming growth factor-alpha (TGF-alpha) in human bone marrow: demonstration of TGF-alpha in erythroblasts and eosinophilic precursor cells and of epidermal growth factor receptors in blastlike cells of myelomonocytic origin. Blood 1995; 85:2385-2392.

30. Anchan RM, Quaas P, Gerami-Naini B, Bartake H, Griffin A, Zhou Y, et al. Amniocytes can serve a dual function as a source of iPS cells and feeder layers. Hum Mol Genet 2011; 20:962-974.

31. Miryounesi M, Nayernia K, Dianatpour M, Mansouri F, Modarressi MH. Co-culture of mouse embryonic stem cells with sertoli cells promote in vitro generation of germ cells. Iran J Basic Med Sci 2013; 16:779-783.

32. Sadat Hashemi Z, Forouzandeh Moghadam M, Soleimani M. Comparison of the ex vivo expansion of UCB-derived CD34+ in 3D DBM/MBA scaffolds with USSC as a feeder layer. Iran J Basic Med Sci 2013; 16:1075-1087.

33. Kim MJ, Shin KS, Jeon JH, Lee DR, Shim SH, Kim JK, et al. Human chorionic-plate-derived mesenchymal stem cells and Wharton's jelly-derived mesenchymal stem cells: a comparative analysis of their potential as placenta-derived stem cells. Cell Tissue Res 2011; 346:53-64.

34. Weiss ML, Anderson C, Medicetty S, Seshareddy KB, Weiss RJ, VanderWerff I, et al. Immune properties of human umbilical cord Wharton's jelly-derived cells. Stem Cells 2008; 26:2865-2874.

35. Qing T, Shi Y, Qin H, Ye X, Wei W, Liu H, et al. Induction of oocyte-like cells from mouse embryonic stem cells by co-culture with ovarian granulosa cells. Differentiation 2007; 75:902-911.

36. Watabe T, Miyazono K. Roles of TGF-beta family signaling in stem cell renewal and differentiation. Cell Res 2009; 19:103-115.

37. Parolini O, Caruso M. Review: Preclinical studies on placenta-derived cells and amniotic membrane: an update. Placenta 2011; 32:S186-195.

38. Kehler J, Tolkunova E, Koschorz B, Pesce M, Gentile L, Boiani M, et al. Oct4 is required for primordial germ cell survival. EMBO Rep 2004; 5:1078-1083.

39. Okamura D, Tokitake Y, Niwa H, Matsui Y. Requirement of Oct3/4 function for germ cell specification. Dev Biol 2008; 317:576-584.

40. Kerr CL, Gearhart JD, Elliott AM, Donovan PJ. Embryonic germ cells: when germ cells become stem cells. Semin Reprod Med 2006; 24:304-313.

41. Berube K, Pitt A, Hayden P, Prytherch Z, Job C. Filter-well technology for advanced three-dimensional cell culture: perspectives for respiratory research. Altern Lab Anim 2010; 38:49-65.