Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Analysis of Apoptosis in Cultured Human Vitrified Ovarian Tissue in the Presence of Leukemia Inhibitory Factor

Authors
Maasoume Abdollahi 1
Mojdeh Salehnia 1
Saghar Salehpour 2
Shahram Pour Beiranvand 1
1 Department of Anatomical Sciences, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran
2 Infertility and Reproductive Health Research Center (IRHRC), Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract
Background: For improving the human ovarian tissue culture, this study was designed to assess the incidence of apoptosis in this tissue following vitrification and in vitro culture in the presence of leukemia inhibitory factor (LIF) as an anti-apoptotic factor. Methods: After collecting the ovarian tissue samples they were divided into non-vitrified and vitrified groups and cultured for 14 days in the presence and absence of LIF then morphological, ultrastructural and steroidogenesis studies, TUNEL and caspase-3/7 assays, and apoptosis analysis by real time RT-PCR were done in all groups. The data were analyzed by independent t-tests and the real time RT-PCR results were compared by one-way ANOVA (p-values of <0.05 were considered significant). Results: No significant difference was observed between non-vitrified and vitrified groups in normality rate of follicles, the levels of hormones, TUNEL positive cells and caspase-3/7 activity. But in all LIF-treated groups, the levels of 17-β estradiol and progesterone were higher and TUNEL signals and caspase-3/7 activity were lower than non-LIF treated groups. The expression of Fas and FasL genes was higher in vitrified group in comparison with non-vitrified group but the expression of other genes was not significantly different. In LIF- treated groups, the expression of pro-apoptotic genes was significantly lower and the expression of anti-apoptotic genes was higher than non-LIF treated group. Conclusion: The vitrification of human ovarian tissue did not increase the incidence of apoptosis at the morphological and molecular levels during long term culture and LIF improves the survival and development of cultured follicles.
Keywords
in vitro culture
Apoptosis
caspase
3/7
Leukemia Inhibitory Factor
Ovarian tissue
Vitrification
  1. Chaput L, Grémeau AS, Vorilhon S, Pons H, Chabrot C, Grèze V, et al. [Fertility preservation in oncology]. Bull Cancer. 2018;105(1):99-110. French.
  2. Shi Q, Xie Y, Wang Y, Li S. Vitrification versus slow freezing for human ovarian tissue cryopreservation: a systematic review and meta-anlaysis. Sci Rep. 2017;7(1):8538.
  3. Xiao Z, Zhang Y, Fan W. Cryopreservation of human ovarian tissue using the silver closed vitrification system. J Assist Reprod Genet. 2017;34(11):1435-44.
  4. Ladanyi C, Mor A, Christianson MS, Dhillon N, Segars JH. Recent advances in the field of ovarian tissue cryopreservation and opportunities for research. J Assist Reprod Genet. 2017;34(6):709-22.
  5. Guzy L, Demeestere I. Assessment of ovarian reserve and fertility preservation strategies in children treated for cancer. Minerva Ginecol. 2017;69(1):57-67.
  6. Zhou XH, Zhang D, Shi J, Wu YJ. Comparison of vitrification and conventional slow freezing for cryopreservation of ovarian tissue with respect to the number of intact primordial follicles: a meta-analysis. Medicine (Baltimore). 2016;95(39):e4095.
  7. Ramezani M, Salehnia M, Jafarabadi M. Short term culture of vitrified human ovarian cortical tissue to assess the cryopreservation outcome: Molecular and morphological analysis. J Reprod Infertil. 2017;18(1):162-71.
  8. Shams Mofarahe Z, Salehnia M, Ghaffari Novin M, Ghorbanmehr N, Fesharaki MG. Expression of folliculogenesis-related genes in vitrified human ovarian tissue after two weeks in vitro culture. Cell J. 2017;19(1):18-26.
  9. Carlsson IB, Laitinen MP, Scott JE, Louhio H, Velentzis L, Tuuri T, et al. Kit ligand and c-Kit are expressed during early human ovarian follicular development and their interaction is required for the survival of follicles in long-term culture. Reproduction. 2006;131(4):641-9.
  10. Telfer EE, McLaughlin M, Ding C, Thong KJ. A two-step serum-free culture system supports development of human oocytes from primordial follicles in the presence of activin. Hum Reprod. 2008;23(5):1151-8.
  11. Arici A, Oral E, Bahtiyar O, Engin O, Seli E, Jones EE. Leukaemia inhibitory factor expression in human follicular fluid and ovarian cells. Hum Reprod. 1997;12(6):1233-9.
  12. Moon C, Liu BQ, Kim SY, Kim EJ, Park YJ, Yoo JY, et al. Leukemia inhibitory factor promotes olfactory sensory neuronal survival via phosphoinositide 3‐kinase pathway activation and Bcl‐2. J Neurosci Res. 2009;87(5):1098-106.
  13. Hunt L, Tudor E, White J. Leukemia inhibitory factor-dependent increase in myoblast cell number is associated with phosphotidylinositol 3-kinase-mediated inhibition of apoptosis and not mitosis. Exp Cell Res. 2010;316(6):1002-9.
  14. Xu J, Li Z, Xu P, Yang Z. Protective effects of leukemia inhibitory factor against oxidative stress during high glucose-induced apoptosis in podocytes. Cell Stress Chaperones. 2012;17(4):485-93.
  15. Furue M, Okamoto T, Hayashi Y, Okochi H, Fujimoto M, Myoishi Y, et al. Leukemia inhibitory factor as an anti-apoptotic mitogen for pluripotent mouse embryonic stem cells in a serum-free medium without feeder cells. In Vitro Cell Dev Biol Anim. 2005;41(1-2):19-28.
  16. Slaets H, Dumont D, Vanderlocht J, Noben JP, Leprince P, Robben J, et al. Leukemia inhibitory factor induces an antiapoptotic response in oligodendrocytes through Akt‐phosphorylation and up‐regulation of 14‐3‐3. Proteomics. 2008;8(6):1237-47.
  17. Azari MF, Profyris C, Karnezis T, Bernard CC, Small DH, Cheema SS, et al. Leukemia inhibitory factor arrests oligodendrocyte death and demyelination in spinal cord injury. J Neuropathol Exp Neurol. 2006;65(9):914-29.
  18. Jo C, Kim H, Jo I, Choi I, Jung SC, Kim J, et al. Leukemia inhibitory factor blocks early differentiation of skeletal muscle cells by activating ERK. Biochim Biophys Acta Mol Cell Res. 2005;1743(3):187-97.
  19. Majumder A, Banerjee S, Harrill JA, Machacek DW, Mohamad O, Bacanamwo M, et al. Neurotrophic effects of leukemia inhibitory factor on neural cells derived from human embryonic stem cells. Stem Cells. 2012;30(11):2387-99.
  20. Nilsson EE, Kezele P, Skinner MK. Leukemia inhibitory factor (LIF) promotes the primordial to primary follicle transition in rat ovaries. Mol Cell Endocrinol. 2002;188(1-2):65-73.
  21. Haidari K, Salehnia M, Valoujerdi MR. The effects of different concentrations of leukemia inhibitory factor on the development of isolated preantral follicles from fresh and vitrified mouse ovaries. Iran Biomed J. 2006;10(4):185-90.
  22. Zhou XH, Wu YJ, Shi J, Zheng SS. Cryopreservation of human ovarian tissue: Comparison of novel direct cover vitrification and conventional vitrification. Cryobiology. 2010;60(2):101-5.
  23. Chang HJ, Moon JH, Lee JR, Jee BC, Suh CS, Kim SH. Optimal condition of vitrification method for cryopreservation of human ovarian cortical tissues. J Obstet Gynaecol Res. 2011;37(8):1092-101.
  24. Xiao Z, Wang Y, Li L, Luo S, Li SW. Needle immersed vitrification can lower the concentration of cryoprotectant in human ovarian tissue cryopreservation. Fertil Steril. 2010;94(6):2323-8.
  25. Salehnia M, Sheikhi M, Pourbeiranvand S, Lundqvist M. Apoptosis of human ovarian tissue is not increased by either vitrification or rapid cooling. Reprod Biomed Online. 2012;25(5):492-9.
  26. Abdollahi M, Salehnia M, Salehpour S, Ghorbanmehr N. Human ovarian tissue vitrification/warming has minor effect on the expression of apoptosis-related genes. Iran Biomed J. 2013;17(4):179-86.
  27. Salehnia M, Moghadam EA, Velojerdi MR. Ultrastructure of follicles after vitrification of mouse ovarian tissue. Fertil Steril. 2002;78(3):644-5.
  28. Huang L, Mo Y, Wang W, Li Y, Zhang Q, Yang D. Cryopreservation of human ovarian tissue by solid-surface vitrification. Eur J Obstet Gynecol Reprod Biol. 2008;139(2):193-8.
  29. Carrasco RA, Stamm NB, Patel BK. One-step cellular caspase-3/7 assay. Biotechniques. 2003;34(5):1064-7.
  30. Pfaffl MW. Quantification strategies in real-time PCR. In: Bustin SA, editor. AZ of quantitative PCR. USA: International University Line; 2004. p. 89-113.
  31. Gandolfi F, Paffoni A, Papasso Brambilla E, Bonetti S, Brevini TA, Ragni G. Efficiency of equilibrium cooling and vitrification procedures for the cryopreservation of ovarian tissue: comparative analysis between human and animal models. Fertil Steril. 2006;85 Suppl 1:1150-6.
  32. Rahimi G, Isachenko V, Todorov P, Tawadros S, Mallmann P, Nawroth F, et al. Apoptosis in human ovarian tissue after conventional freezing or vitrification and xenotransplantation. Cryo Lett. 2009;30(4):300-9.
  33. Dalman A, Deheshkar Gooneh Farahani NS, Totonchi M, Pirjani R, Ebrahimi B, Rezazadeh Valojerdi M. Slow freezing versus vitrification technique for human ovarian tissue cryopreservation: an evaluation of histological changes, WNT signaling pathway and apoptotic genes expression. Cryobiology. 2017;79:29-36.
  34. Brodowska A, Laszczyńska M, Starczewski A. Apoptosis in ovarian cells in postmenopausal women. Folia Histochem Cytobiol. 2007;45(2):99-105.
  35. Abir R, Fisch B, Jin S, Barnnet M, Freimann S, Van den Hurk R, Feldberg D, et al. Immunocytochemical detection and RT–PCR expression of leukaemia inhibitory factor and its receptor in human fetal and adult ovaries. Mol Hum Reprod. 2004;10(5):313-9.
  36. Da Nóbrega JE, Gonçalves PBD, Chaves RN, Magalhaes DdM, Rossetto R, Lima-Verde IB, et al. Leukemia inhibitory factor stimulates the transition of primordial to primary follicle and supports the goat primordial follicle viability in vitro. Zygote-The Biology of Gametes and Early Embryos. 2012;20(1):73-8.
  37. Haidari K, Salehnia M, Rezazadeh Valojerdi M. The effect of leukemia inhibitory factor and coculture on the in vitro maturation and ultrastructure of vitrified and nonvitrified isolated mouse preantral follicles. Fertil Steril. 2008;90(6):2389-97.
  38. Kong HS, Kim EJ, Youm HW, Kim SK, Lee JR, Suh CS, et al. Improvement in ovarian tissue quality with supplementation of antifreeze protein during warming of vitrified mouse ovarian tissue. Yonsei Med J. 2018;59(2):331-6.
  39. Lalitkumar S, Boggavarapu NR, Menezes J, Dimitriadis E, Zhang JG, Nicola NA, et al. Polyethylene glycated leukemia inhibitory factor antagonist inhibits human blastocyst implantation and triggers apoptosis by down-regulating embryonic AKT. Fertil Steril. 2013;100(4):1160-9.
  40. Townson DH, Combelles CM. Ovarian follicular atresia. Basic gynecology some related issues in tech. 2012:156.
  41. Yang R, Xu S, Zhao Z, Li J. Fas ligand expression and mediated activation of an apoptosis program in bovine follicular granulosa cells. Gene. 2012;493(1):148-54.
  42. Tilly J, Tilly K, Kenton M, Johnson A. Expression of members of the bcl-2 gene family in the immature rat ovary: equine chorionic gonadotropin-mediated inhibition of granulosa cell apoptosis is associated with decreased bax and constitutive bcl-2 and bcl-xlong messenger ribonucleic acid levels. Endocrinology. 1995;136(1):232-41.
  43. Depalo R, Nappi L, Loverro G, Bettocchi S, Caruso ML, Valentini AM, et al. Evidence of apoptosis in human primordial and primary follicles. Hum Reprod. 2003;18(12):2678-82.
  44. Yang E, Korsmeyer SJ. Molecular thanatopsis: a discourse on the BCL2 family and cell death. Blood. 1996;88(2):386-401.
  45. Reed JC. Double identity for proteins of the Bcl-2 family. Nature. 1997;387(6635):773-6.
  46. Tilly JL. Apoptosis and ovarian function. Rev Reprod. 1996;1(3):162-72.
Journal of Reproduction & Infertility
Volume 19, Issue 4
Autumn 2018
Pages 193-202