Volume 6, Issue 5 (7-2008)                   IJRM 2008, 6(5): 209-215 | Back to browse issues page

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Mahmoudi R, Amiri I, Pasbakhsh P, Ragardi Kashani I, Abbasi M, Aboulhasani F, et al . The effects of vitrification on spindle apparatus of in vitro matured germinal vesicle in mice. IJRM 2008; 6 (5) :209-215
URL: http://ijrm.ir/article-1-126-en.html
1- Department of Anatomy, Faculty of Medicine, Yasouj University of Medical Sciences, Yasuj, Iran
2- Department of Anatomy, Faculty of Medicine, Hamedan University of Medical Sciences, Hamedan, Iran
3- Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran
4- Department of Anatomy, Faculty of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran
5- Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran , Sobhania@tums.ac.ir
Abstract:   (3077 Views)
Background: Routine oocytes cryopreservation remained an elusive technique in the wide ranges of available assisted reproductive technologies. The microtubules of oocytes are vulnerable to cryoprotectants and thermal change during cryopreservation.
Objective: The effects of a vitrification protocol were investigated on the spindle and chromosome configurations of mice oocytes cryopreserved at the germinal vesicle stage.
Materials and Methods: Germinal vesicle with cumulus cells were transferred to vitrification solution which was composed of 30% (v/v) ethylene glycol, 18% Ficoll-70 and 0.3 M Sucrose either by single step or in step-wise way. Following vitrification and in vitro maturation (MII), the matured oocytes were immonostained for meiotic spindles and chromosomes, before visualization using fluorescent microscopy.
Results: A statistically significant increase was observed in the survival and maturation rate in step-wise vitrification (88.96% and 71.23% respectively) compared to single step vitrification (70.6% and 62.42% respectively) (p<0.05). Normal spindle morphology after vitrification-thawing in step-wise vitrification group (77.26%) was higher than single step vitrification group (64.24%) but lower than control group (94.75%) (p<0.05).
Conclusion: The results suggest that vitrification with step-wise procedure on mice germinal vesicle oocytes has positive effects on survival and maturation rate and normal spindle configuration compare with single step vitrification procedure.
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Type of Study: Original Article |

References
1. Wennerholm WB. Cryopreservation of embryos and oocytes: obstetric outcome and health in children. Hum Reprod 2000; 15: 18-25. [DOI:10.1093/humrep/15.suppl_5.18]
2. Aono N, Naganuma T, Abe Y, Hara K, Sasada H, Sato E, et al. Successful production of blastocysts following ultrarapid vitrification with step-wise equilibriation of germinal vesicle-stage mouse oocytes. J Reprod Dev 2003; 49: 501-506. [DOI:10.1262/jrd.49.501]
3. Chen SU, Lien YR, Chao KH, Ho HN, Yang YS, Lee TY. Effects of cryopreservation on meiotic spindles of oocytes and its dynamics after thawing: Clinical implications in oocyte freezing. Molecular and Cellular Endocrinology 2003; 202: 101-107. [DOI:10.1016/S0303-7207(03)00070-4]
4. Tucker MJ, Morton PC, Wright G, Sweitzer CL, Massey JB. Clinical application of human egg cryopreservation. Hum Reprod 1998; 13: 3156-3159. [DOI:10.1093/humrep/13.11.3156]
5. Boiso I, Marti M, Santalo J, Ponsa M, Barri PN, Veiga A. A confocal microscopy analysis of the spindle and chromosome configuration of human oocytes cryopreserved at the germinal vesicle and metaphase II stage. Hum Reprod 2002; 17: 1885-1891. [DOI:10.1093/humrep/17.7.1885]
6. Pickering SJ, Braude PR, Johnson MH, Cant A, Currie J. Transient cooling to room temperature can cause irreversible disruption of the meiotic spindle in the human oocyte. Fertil Steril 1990; 54: 102-108. [DOI:10.1016/S0015-0282(16)53644-9]
7. Almeida P, Bolton V. The effect of temperature fluctuations on the cytoskeletal organisation and chromosomal constitution of the human oocyte. Zygote 1995; 3: 357-365. [DOI:10.1017/S0967199400002793]
8. Van Blerkom J, Davis PW. Cytogenetic, cellular, and developmental consequences of cryopreservation of immature and mature mouse and human oocytes. Microsc Res Tech 1994; 27: 165-193. [DOI:10.1002/jemt.1070270209]
9. Joly C, Bchini O, Boulekbache H. Effects of 1,2-propanediol on the cytoskeletal organization of the mouse oocyte. Hum Reprod 1992; 7: 374-378. [DOI:10.1093/oxfordjournals.humrep.a137654]
10. Eroglu A, Toth TL, Toner M. Alterations of the cytoskeleton and polyploidy induced by cryopreservation of metaphase II mouse oocytes. Fertil Steril 1998; 69: 944-957. [DOI:10.1016/S0015-0282(98)00030-2]
11. Wu J, Zhang L, Wang X. In vitro maturation, fertilization and embryo development after ultrarapid freezing of immature human oocytes. Reproduction 2001; 121: 389-393. [DOI:10.1530/rep.0.1210389]
12. Oktay K, Demirtas E, Son WY, Lostritto K, Chian RC, Tan SL. In vitro maturation of germinal vesicle oocytes recovered after premature luteinizing hormone surge: description of a novel approach to fertility preservation. Fertil Steril. 2008; 89: e19-22. [DOI:10.1016/j.fertnstert.2007.02.028]
13. Kagawa N, Kuwayama M, Nakata K, Vajta G, Silber S, Manabe N, et al. Production of the first offspring from oocytes derived from fresh and cryopreserved pre-antral follicles of adult mice. Reprod Biomed Online 2007; 14: 693-699. [DOI:10.1016/S1472-6483(10)60670-0]
14. Chian RC, Gilbert L, Huang JY, Demirtas E, Holzer H, Benjamin A, et al. Live birth after vitrification of in vitro matured human oocytes. Fertil Steril 2008; 29: 1-5.
15. kassai M, Komi JH, Takakamo A, Tsudera H, Sakurai T. A simple method for mouse embryo cryopreservation in low toxicity vitrification solution, without appreciable loss of viability. J Reprod Fertil 1999; 89: 91-97. [DOI:10.1530/jrf.0.0890091]
16. Johnson MH, Pickering SJ. The effect of dimethylsulphoxide on the microtubular system of mouse oocytes. Development 1987; 100: 313-324.
17. Hong SW, Chung HM, Lim JM, Ko JJ, Yoon TK, Yee B. Improved human oocyte development after vitrification: a comparison of thawing methods. Fertil Steril 1999; 72: 142-146. [DOI:10.1016/S0015-0282(99)00199-5]
18. Yoon TK, Kim TJ, Park SE, Hong SW, Ko JJ, Chung HM. Live births after vitrification of oocytes in a stimulated in vitro fertilizationembryo transfer program. Fertil Steril 2003; 79: 1323-1326. [DOI:10.1016/S0015-0282(03)00258-9]
19. Toth TL, Baka SG, Veek LL, Jones HW, Muasher S, Lanzenddorf SE. Fertilization and in vitro development of cryopreserved human prophase I oocytes.Fertil Steril 1994; 61: 891-894. [DOI:10.1016/S0015-0282(16)56702-8]
20. Hurt AE, Landim F, Seidel GE, Squires EL. Vitrification of immature and mature equine and bovine oocytes in an ethylene glycol, ficoll and sucrose solution using open-pulled straws. Theriogenology 2000; 54: 119-128. [DOI:10.1016/S0093-691X(00)00330-7]
21. Cetin Y, Bastan A. Cryopreservation of immature bovine oocytes by vitrification in straws. Anim Reprod Sci 2006; 92: 29-36. [DOI:10.1016/j.anireprosci.2005.05.016]
22. Aono N, Abe Y, Hara K, Sasada H, Sato E,Yoshida H. Production of live offspring from mouse germinal vesicle-stage oocytes vitrified by a modified stepwise method, SWEID. Fertil Steril 2005; 84: 1078-1082. [DOI:10.1016/j.fertnstert.2005.03.077]
23. Kuwayama M, Fujikawa S, Nagai T. Ultrastructure of IVM-IVF bovine blastocysts vitrified after equilibration in glycerol 1, 2-propanediol using 2-step and 16-step procedures. Cryobiology 1994; 31: 415-422. [DOI:10.1006/cryo.1994.1051]
24. Saunders K, Parks JE. Effects of cryopreservation procedures on the cytology and fertilization rate of in vitro matured bovine oocytes. Biol Reprod 1999; 61: 178-187. [DOI:10.1095/biolreprod61.1.178]
25. Webb M, Howlet S, Maro B. Parthenogenesis and cytoskeletal organization in aging mouse eggs. J Embryol Exp Morphol 1986; 95: 131-145.
26. Magistrini M, Szöllösi D. Effects of cold and of isopropyl-N-phenylcarbamate on the second meiotic spindle of mouse oocytes. Eur J Cell Biol 1980; 22: 699-707.
27. Liu RH, Sun QY, Li YH, Jiao LH, Wang WH. Effects of cooling on meiotic spindle structure and chromosome alignment within in vitro matured porcine oocytes. Mol Reprod 2003; 65: 212-218. [DOI:10.1002/mrd.10282]
28. Wu C, Rui R, Dai J, Zhang C, Ju S, Xie B, et al. Effects of cryopreservation on the developmental competence, ultrastructure and cytoskeletal structure of porcine oocytes. Mol Reprod Dev 2006; 73: 1454-1462. [DOI:10.1002/mrd.20579]
29. Albarracin JL, Morato R, Rojas C, Mogas T. Effects of vitrification in open pulled straws on the cytology of in vitro matured prepubertal and adult bovine oocytes. Theriogenology 2005; 63: 890-901. [DOI:10.1016/j.theriogenology.2004.05.010]

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