The Effect of The Conditioned Medium from Human Embryonic Stem Cells on Mouse Oocytes In Vitro Maturation

Document Type : Original Article

Authors

1 Department of Biology, Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran

2 Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

3 Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

4 Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

Abstract

Objective: Some reports have indicated that conditioned medium from growing mouse embryonic stem cells (ESCs)
provides a supportive condition for small follicles growing, oocyte maturation, and following embryo growth. The aim of
this study is assessing in vitro maturation (IVM) and consequent in vitro fertilization (IVF) outcome of immature mouse
oocytes using human embryonic stem cells conditioned medium (HESCM).
Materials and Methods: In this experimental study, 240 germinal vesicle (GV) oocytes were took from NMRI female
mice, aged 4-6 weeks, 48 hours before injection of 5 IU pregnant mare serum gonadotropin (PMSG). 120 GV oocytes
without cumulus cells were cultured in each of the groups. 120 GV were cultured in HESCM as test groups and also
120 GV cultured in human embryonic stem cells medium (HESM) as control groups. After evaluating the metaphase II
(MII) oocyte maturation rate at 8, 16 and 24 hours, the MII oocytes subsequently were fertilized in vitro and the two-cell
embryo development rate was recorded at days 1, 2, and 3. Statistical analysis was performed by using the generalized
estimating equations (GEE) method that calculated their rate ratio.
Results: Our data indicated there are significant differences between the maturation rates in HESCM and HESM
(P=0.004), also the two-cell embryo development was significant between two culture media (P=0.00).
Conclusion: Similar to some other studies, the secretome of the HESCM showed a significant impact on the IVM
outcomes in mice.

Keywords

Main Subjects


  1. Pincus G, Enzmann EV. The comparative behavior of mammalian eggs in vivo and in vitro: I. The activation of ovarian eggs. J Exp Med. 1935; 62(5): 665-675.
  2. Dahan MH, Tan SL, Chung J, Son WY. Clinical definition paper on in vitro maturation of human oocytes. Hum Reprod. 2019; 31(7): 1383-1386.
  3. De Vos M, Smitz J, Thompson JG, Gilchrist RB. The definition of IVM is clear-variations need defining. Hum Reprod. 2019; 31(11):2411-2415.
  4. Sanchez F, Lolicato F, Romero S, DeVos M, Van Ranst H, Verheyen G, et al. An improved IVM method for cumulus oocyte complexes from small follicles in polycystic ovary syndrome patients enhances oocyte competence and embryo yield. Hum Reprod. 2017; 32(10): 2056-2068.
  5. Saenz-de-Juano MD, Ivanova E, Romero S, Lolicato F, Sanchez F, Van Ranst H, et al. DNA methylation and mRNA expression of imprinted genes in blastocysts derived from an improved in vitro maturation method for oocytes from small antral follicles in polycystic ovary syndrome patients. Hum Reprod. 2019; 34(9): 1640-1649.
  6. Yang ZY, Chian RC. Development of in vitro maturation techniques for clinical applications. Fertil Steril. 2017; 108(4): 577-584.
  7. Adib M, Seifati SM, Dehghani Ashkezari M, Akyash F, Khoradmehr A, Aflatoonian B. Effect of human testicular cells conditioned medium on in vitro maturation and morphology of mouse oocytes. Int J Fertil Steril. 2020; 14(3): 175-184.
  8. Adib M, Seifati SM, Ashkezari MD, Khoradmehr A, Rezaee-Ranjbar- Sardari R, Tahajjodi SS, et al. The effect of the human cumulus cells-conditioned medium on in vitro maturation of mouse oocyte. Int J Reprod BioMed. 2020; 18(12): 1019-1028.
  9. Akyash F, Sadeghian-Nodoushan F, Tahajjodi SS, Nikukar H, Farashahi Yazd E, Azimzadeh M, et al. Human embryonic stem cells and good manufacturing practice: report of a 1-day workshop held at stem cell biology research center. Int J Reprod Biomed. 2017; 15(5): 255-256.
  10. Akyash F, Tahajjodi S, Farashahi Yazd E, Hajizadeh-Tafti F, Sadeghian- Nodoushan F, Golzadeh J, et al. Derivation of new human embryonic stem cell lines (Yazd1-3) and their vitrification using cryotech and cryowin tools: a lab resources report. Int J Reprod Biomed. 2019; 17(12): 891-906.
    11. Fatma S, Selby DE, Singla RD, Singla DK. Factors released from embryonic stem cells stimulate c-kit-FLK1 (+ve) progenitor cells and enhance neovascularization. Antioxid Redox Signal. 2010; 13(12): 1857-1865.
    12. Miraki S, Mokarizadeh A, Banafshi O, Assadollahi V, Abdi M, Roshani D, et al. Embryonic stem cell conditioned medium supports in vitro maturation of mouse oocytes. Avicenna J Med Biotech. 2017; 9(3): 114-119.
    13. Richani D, Gilchrist RB. The epidermal growth factor network: role in oocyte growth, maturation and developmental competence. Hum Reprod Update. 2018; 24(1): 1-14.
    14. Yeo CX, Gilchrist RB, Thompson JG, Lane M. Exogenous growth differentiation factor 9 in oocyte maturation media enhances subsequent embryo development and fetal viability in mice. Hum Reprod. 2008; 23(1): 67-73.
    15. Hussein TS, Sutton-McDowall ML, Gilchrist RB, Thompson JG. Temporal effects of exogenous oocyte-secreted factors on bovine oocyte developmental competence during IVM. Reprod Fertil Dev. 2011; 23(4): 576-584.
    16. Sutton-McDowall ML, Mottershead DG, Gardner DK, Gilchrist RB, Thompson JG. Metabolic differences in bovine cumulus-oocyte complexes matured in vitro in the presence or absence of folliclestimulating hormone and bone morphogenetic protein 15. Biol Reprod. 2012; 87(4): 87.
    17. Son WY, Henderson S, Cohen Y, Dahan M, Buckett W. Immature oocyte for fertility preservation. Front Endocrinol (Lausanne). 2019; 10: 464.
    18. Pawshe CH, Appa Rao KB, Jain SK, Totey SM. Biochemical studies on goat oocytes: Timing of nuclear progression, effect of protein inhibitor and pattern of polypeptide synthesis during in vitro maturation. Theriogenology. 1994; 42(2): 307-320.
    19. Giuffrida D, Rogers IM, Nagy A, Calogero AE, Brown TJ, Casper RF. Human embryonic stem cells secrete soluble factors that inhibit cancer cell growth. Cell Prolif. 2009; 42(6): 788-798.
    20. Chian RC, Tan SL. Maturational and developmental competence of cumulus-free immature human oocytes derived from stimulated and intracytoplasmic sperm injection cycles. Reprod Biomed Online. 2002; 5(2): 125-132.
    21. Farsi MM, Jorsaraei SG, Esmaelzadeh S, Golaipour MJ. In vitro maturation of germinal vesicle oocytes in stimulated intracytoplasmic sperm injection cycles. Cell J. 2011; 13(2): 73-78.
    22. Kim BK, Lee SC, Kim KJ, Han CH, Kim JH. In vitro maturation, fertilization, and development of human germinal vesicle oocytes collected from stimulated cycles. Fertil Steril. 2000; 74(6): 1153-1158.
    23. Lee SR, Kim MO, Kim SH, Kim BS, Yoo DH, Park YS, et al. Effect of conditioned medium of mouse embryonic fibroblasts produced from EC-SOD transgenic mice in nuclear maturation of canine oocytes in vitro. Anim Reprod Sci. 2007; 99(1-2): 106-116.
    24. Abdel-Ghani MA, Abe Y, Asano T, Hamano S, Suzuki H. Effect of bovine cumulus-oocyte complexes-conditioned medium on in vitro maturation of canine oocytes. Reprod Med Biol. 2010; 26; 10(1): 43-49.
    25. Izadi M, Rezvani ME, Aliabadi A, Karimi M, Aflatoonian B. Mesenchymal stem cells-derived exosomes as a promising new approach for the treatment of infertility caused by polycystic ovary syndrome. Front Pharmacol. 2022; 13: 1021581.