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Skip Navigation LinksJRI > Archive > January-March 2004, Volume 5, Issue 1 > Effect of vitrification on apoptosis in mouse blastocysts



Volume 5, Issue 1, Number 17 / January-March
(pages 14-22)


Effect of vitrification on apoptosis in mouse blastocysts




 Corresponding Author
Department of Anatomy Sciences, Faculty of Medicine, Ghazvin University of Medical Sciences, Ghazvin, Iran

Anatomy Department, Faculty of Medicine, Tabriz Medical Sciences University, Tabriz, Iran

Laboratory of Histology & Cellular Biology, Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

Department of Statistic & Epidemiology Faculty of Health & Nutrition, Tabriz University of Medical Sciences, Tabriz, Iran


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Abstract
Introduction: In recent years there have been a great advances in vitrification of embryos. However, there is no reliable vitrification protocol to ensure a high embryo survival rate, Because the mechanisms of embryo injury has not been discovered precisely. The aim of the present study was to determine the effects of vitrification on apoptosis in mouse blastocysts. Materials and Methods: Ninety five mouse blastocysts were obtained by flushing from Swiss Albino mouse and randomly divided into control and experimental groups. Blastocysts in the control group (52) were cultured in M16 media for 2 hours and then the apoptotic index were obtained after staining by TUNEL technique with PI. Blastocysts in the experimental group (43) were vitrified just after flushing in EFS40 solution and kept in LN2 for one month. After thawing and culture in M16 for 2 hours, the apoptotic indices were obtained by TUNEL staining. Results: The results showed that the mean number of blastomers in the vitrified blastocysts group (44.912.47) was not significantly different (P=0.176) from those that seen in the control group (50.232.9), while the mean number of apoptotic blastomers in vitrified blastocysts group (4.080.28) was significantly higher (P=0.02) as compared to the control group (4.930.22). The mean apoptosis Index in vitrified blastocysts (11.870.63) was significantly higher (P<0.004) than the control group (9.120.67). Conclusion: we can conclude that the vitrification can increase apoptotic cell death in mouse blastocysts.

Keywords: Vitrification, Apoptosis, Mouse blastocyst, TUNEL


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References
  1. Kasai M. Cryopreservation of mammalian embryos. Mol Biotechnol.1997;7(2):173-9.
  2. Dobrinsky J.R., Pursel V.G., Long C.R., Johnson L.A. Birth of piglets after transfer of embryos cryopreserved by cytoskeletal stabili- zation and vitrification. Biol Reprod.2000;62(3): 564-70.
  3. Kasai M., Ito K., Edashige K. Morphological appearance of the cryopreserved mouse blastocyst as a tool to identify the type of cryoinjury. Hum Reprod.2002;17(7):1863-74.
  4. Kasai M., Zhu S.E., Pedro P.B., Nakamura K., Sakurai T., Edashige K. Fracture damage of embryos and its prevention during vitrification and warming. Cryobiology.1996;33(4):459-64.
  5. Whittingham D.G., Leibo S.P., Mazur P. Survival of mouse embryos frozen to–196°C and –269°C. Science.1972;178:411-414.
  6. Rall W.F., Fahy G.M. Ice-free cryopreservation of mouse embryos at-196 degrees C by vitrification. Nature.1985;313(6003):573-5.
  7. Gavrieli Y., Sherman Y., Ben- Sasson S.A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation.J Cell Biol.1992;119(3):493-501.
  8. Hardy K. Cell death in the mammalian blastocyst. Mol Hum Reprod.1997;3:919– 92.
  9. Gavrieli Y., Sherman Y., Ben-Sasson S.A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol.1992;119(3):493-501.
  10. Yamadori I., Yoshino T., Kondo E., Cao L., Akagi T., Matsuo Y., Minowada J. Comparison of two methods of staining apoptotic cells of transferase and DNA polymerase I reactions. J leukemia cell lines. Terminal deoxynucleotidyl Histochem Cytochem.1998;46(1):85-90.
  11. Neuber E., Luetjens C.M., Chan A.W., Schatten G.P. Analysis of DNA fragmentation of in vitro cultured bovine blastocysts using TUNEL. Theriogenology.2002;57(9):2193-202.
  12. Pampfer S., Vanderheyden I., McCracken J.E., Vesela J., Hertogh R. Increased cell death in rat blastocysts exposed to maternal diabetes in utero and to high glucose or tumor necrosis factor-alpha in vitro. Development.1997;124(23): 4827-36.
  13. Whittingham D.G. Survival of mouse embryos after freezing and thawing.Nature. 1971.10;233(5315):125-6.
  14. Edashige K., Asano A., An T.Z., Kasai M. Restoration of resistance to osmotic swelling of vitrified mouse embryos by short-term culture. Cryobiology.1999;38(4):273-80.
  15. Zhu S.E., Kasai M., Otoge H., Sakurai T., Machida T. Cryopreservation of expanded mouse blastocysts by vitrification in ethylene glycol-based solutions. J Reprod Fertil.1993;98(1):139-45.
  16. Brison DR, Schultz RM. Apoptosis during mouse blastocyst formation: evidence for a role for survival factors including transforming growth factor alpha. Biol Reprod 1997:56:1088-1096.
  17. Yuge M, Otoi T, Nii M, Murakami M, Karja NW, Rajaei F, Agung B, wongsrikeao P, Murakami M, Suzuki T. Effects of cooling ovaries before oocyte aspiration on meiotic competence of porcine oocytes and of exposing in vitro matured oocytes to ambient temperature on in vitro fertilitzation and development of the oocytes. Cryobiology.2003 Oct;47(2):102-8.
  18. Munne s, alikani M, Tomkin G, Grifo J, Cohen J. Embryo morphology, developmental rates, and maternal age are correlated with chromosome abnormalities. Fertil Steril.1995;64(2):382-91.
  19. Hovatta O. Cryopreservation and culture of human primordial and primary ovarian follicles. Mol Cell Endocrinol.2000;169(1-2):95-7.
  20. Rahimi G., Isachenko E., Sauer H., Isachenko V., Wartenberg M., Hescheler J., Mallmann P. Nawroth F Effect of different vitrification protocols for human ovarian tissue on reactive oxygen species and apoptosis. Reprod Fertil Dev.2003;15(6):343-9.
  21. Demirei B, Salle B, Frappart L, Franck M, Guerin JF, Lornage J. Morphological alterations and DNA fragmentation in oocytes from primordial and primary follicles after freezing- thawing of ovarian cortex in sheep. Fertil Steril. 2003;77(3):595-600.
  22. Niemann H., Lucas- Hahn A., Stoffregen C. Cryopreservation of bovine oocytes and embryos following microsurgical operations. Mol Reprod Dev.1993;36(2):232-5.
  23. Bernard A., Fuller B.J. Cryopreservation of human oocytes: a review of current problems and perspectives. Hum Reprod Update.1996;2(3):193-207.
  24. Kola I., Kirby C., Shaw J., Davey A., Trounson A. Vitrification of mouse oocytes results in aneuploid zygotes and malformed fetuses. Teratology.1988;38(5):467-74.
  25. Vincent C, Johnson MH. Cooling, cryo- prodtectants, and the cytoskelieton of the mammalian oocyte. Oxf Rev Reprod Biol.1992; 14:73-100.
  26. Lane M., Lyons E.A., Bavister B.D. Cryo-preservation reduces the ability of hamster 2-cell embryos to regulate intracellular pH. Hum Reprod 2000;15(2):389-94.
  27. Yang M.Y., Rajamahendran R. Expression of Bcl-2 and Bax proteins in relation to quality of bovine oocytes and embryos produced in vitro. Anim Reprod Sci.2002;70(3-4):159-69.



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