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Effect of Cytochalasin B Treatment on the Improvement of Survival Rate in Vitrified Pig Oocyte

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초록

To improve survival rates of vitrified pig oocytes, the treatment of cytoskeletal stabilizer on an appropriate time is one of the possible approaches. However, the exact treatment timing and effect of cytoskeletal stabilizer such as cyto-chalasin B (CB) is not well known during oocyte vitrification procedures. Thus, the present study was conducted to determine optimal treatment timing of CB during vitrification and warming procedures. In experiment 1, the survival rates of the post-warming pig oocytes were analyzed by fluorescein diacetate (FDA) assays with 4 classifications. In results, post-warming oo-cytes showed significantly (p<0.05) decreased number of alive oocytes (31.8% vs. 86.4%) compared to fresh control. In detail, the significant difference (p<0.05) was found only in strong fluorescence (18.2% vs. 70.5%) not in intermediate fluorescence groups (13.6% vs. 15.9%). In experiment 2, CB was treated before (CB-Vitri) and after (Vitri-CB) vitrification. In results, group of Vitri-CB showed significantly (p<0.05) higher (91.6%) survival rates compared to group of CB-Vitri (83.7%), sig-nificantly (p<0.05) and comparable with group of Vitri Control (88.7%) by morphological inspection. In FDA assay results, group of Vitri-CB showed significantly (p<0.05) higher (44.2%) survival rates compared to groups of CB-Vitri (36.7%) and Vitri Control (35.1%). In conclusion, the increased survival rates of post-warming pig oocyte treated with Vitri-CB method are firstly described here. The main finding of present study is that the CB treatment during recovery could be helpful to re-fresh the post-warming pig oocyte resulting its improved survival rates.

Abstract

To improve survival rates of vitrified pig oocytes, the treatment of cytoskeletal stabilizer on an appropriate time is one of the possible approaches. However, the exact treatment timing and effect of cytoskeletal stabilizer such as cyto-chalasin B (CB) is not well known during oocyte vitrification procedures. Thus, the present study was conducted to determine optimal treatment timing of CB during vitrification and warming procedures. In experiment 1, the survival rates of the post-warming pig oocytes were analyzed by fluorescein diacetate (FDA) assays with 4 classifications. In results, post-warming oo-cytes showed significantly (p<0.05) decreased number of alive oocytes (31.8% vs. 86.4%) compared to fresh control. In detail, the significant difference (p<0.05) was found only in strong fluorescence (18.2% vs. 70.5%) not in intermediate fluorescence groups (13.6% vs. 15.9%). In experiment 2, CB was treated before (CB-Vitri) and after (Vitri-CB) vitrification. In results, group of Vitri-CB showed significantly (p<0.05) higher (91.6%) survival rates compared to group of CB-Vitri (83.7%), sig-nificantly (p<0.05) and comparable with group of Vitri Control (88.7%) by morphological inspection. In FDA assay results, group of Vitri-CB showed significantly (p<0.05) higher (44.2%) survival rates compared to groups of CB-Vitri (36.7%) and Vitri Control (35.1%). In conclusion, the increased survival rates of post-warming pig oocyte treated with Vitri-CB method are firstly described here. The main finding of present study is that the CB treatment during recovery could be helpful to re-fresh the post-warming pig oocyte resulting its improved survival rates.

참고문헌 (34)

  1. Albarracin JL, "Effects of vitrification in open pulled straws on the cytology of in vitro matured prepubertal and adult bovine oocytes", Theriogenology, 63, pp.890~901 (2005)
  2. Almasi Turk S, "Mouse oocytes and embryos cryotop-vitrification using low concentrated solutions: Effects on meiotic spindle, genetic material array and developmental ability", Iran J Basic Med Sci, 16, pp.599~601 (2013)
  3. Amoushahi M, "Vitrification of mouse MII oocyte decreases the mitochondrial DNA copy number, TFAM gene expression and mitochondrial enzyme activity", J Reprod Infertil, 18, pp.343~351 (2017)
  4. Balaban B, "Practical Manual of In Vitro Fertilization: Advanced Methods and Novel Devices" (2012), Springer
  5. Balaban B, "Oocyte morphology does not affect fertilization rate, embryo quality and implantation rate after intracytoplasmic sperm injection", Hum Reprod, 13, pp.3431~3433 (1998)
  6. Boyer P, "Oocyte vitrification and its impact on the clinical practice of assisted reproduction", Gynecol Obstet Fertil, 40, pp.687~690 (2012)
  7. Chasombat J, "Pretreatment of in vitro matured bovine oocytes with docetaxel before vitrification: Effects on cytoskeleton integrity and developmental ability after warming", Cryobiology, 71, pp.216~223 (2015)
  8. Coticchio G, "Vitrification may increase the rate of chromosome misalignment in the metaphase II spindle of human mature oocytes", Reprod Biomed Online, 3, pp.29~34 (2009)
  9. Dai J, "Changes in mitochondrial function in porcine vitrified MII-stage oocytes and their impacts on apoptosis and developmental ability", Cryobiology, 41, pp.291~298 (2015)
  10. De Sutter P, "Oocyte morphology does not correlate with fertilization rate and embryo quality after intracytoplasmic sperm injection", Hum Reprod, 11, pp.595~597 (1996)
  11. dela Pena EC, "Vitrification of mouse oocytes in ethylene glycol-raffinose solution: Effects of preexposure to ethylene glycol or raffinose on oocyte viability", Cryobiology, 42, pp.103~111 (2001)
  12. Didion BA, "Observations on the cooling and cryopreservation of pig oocytes at the germinal vesicle stage", J Anim Sci, 68, pp.2803~2810 (1990)
  13. Fabbri R, "Oocyte cryopreservation", Hum Reprod, 13, pp.98~108 (1998)
  14. Fuku E, "Ultrastructural changes in bovine oocytes cryopreserved by vitrification", Cryobiology, 32, pp.139~156 (1995)
  15. Hamamah S, "Oocyte and embryo quality: Is their morphology a good criterion?", J Gynecol Obstet Biol Reprod (Paris), 34, pp.5S38~5S41 (2005)
  16. Hara H, "High incidence of multiple aster formation in vitrified-warmed bovine oocytes after invitro fertilization", Theriogenology, 77, pp.908~915 (2012)
  17. Hwang IS, "Rescue of vitrified-warmed bovine oocytes with rho-associated coiled-coil kinase inhibitor", Biol Reprod, 89, pp.1~6 (2013)
  18. Hwang IS, "Recent progress in cryopreservation of bovine oocytes", Biomed Res Int, 2014, pp.570647~ (2014)
  19. Hwang IS, "Improved survival and developmental rates in vitrified-warmed pig oocytes after recovery culture with coenzyme Q10", Cryo Letters, 37, pp.59~67 (2016)
  20. 황인설, "Production of Cloned Korean Native Pig by Somatic Cell Nuclear Transfer", 발생과 생식, 19(2), pp.79~84 (2015), 한국발생생물학회
  21. Isachenko V, "Vitrification of immature porcine oocytes: Effects of lipid droplets, temperature, cytoskeleton, and addition and removal of cryoprotectant", Cryobiology, 36, pp.250~253 (1998)
  22. Kikuchi K, "Morphological features of lipid droplet transition during porcine oocyte fertilisation and early embryonic development to blastocyst in vivo and in vitro", Zygote, 10, pp.355~366 (2002)
  23. Kono T, "Development of vitrified mouse oocytes after in vitro fertilization", Cryobiology, 28, pp.50~54 (1991)
  24. Lowther KM, "Maturation, fertilization, and the structure and function of the endoplasmic reticulum in cryopreserved mouse oocytes", Biol Reprod, 81, pp.147~154 (2009)
  25. Marco-Jimenez F, "Effect if cytochalasin B pre-treatment of in vitro matured porcine oocytes before vitrification", Cryo Letters, 33, pp.24~30 (2012)
  26. Mohr LR, "The use of fluorescein diacetate to assess embryo viability in the mouse", J Reprod Fertil, 58, pp.189~196 (1980)
  27. Mullen SF, "A chronologic review of mature oocyte vitrification research in cattle, pigs, and sheep", Theriogenology, 78, pp.1709~1719 (2012)
  28. Noto V, "Fluorescein diacetate assessment of embryo viability after ultrarapid freezing of human multipronucleate embryos", Fertil Steril, 55, pp.1171~1175 (1991)
  29. Pitchayapipatkul J, "Microtubule stabilisers docetaxel and paclitaxel reduce spindle damage and maintain the developmental competence of in vitro-mature bovine oocytes during vitrification", Reprod Fertil Dev, 29, pp.2028~2039 (2017)
  30. Rojas C, "Vitrification of immature and in vitro matured pig oocytes: Study of distribution of chromosomes, microtubules, and actin microfilaments", Cryobiology, 49, pp.211~220 (2004)
  31. Somfai T, "Factors affecting cryopreservation of porcine oocytes", J Reprod Dev, 58, pp.17~24 (2012)
  32. Vallorani C, "Pig oocyte vitrification by Cryotop method and the activation of the apoptotic cascade", Anim Reprod Sci, 135, pp.68~74 (2012)
  33. Van Soom A, "Assessment of mammalian embryo quality: What can we learn from embryo morphology?", Reprod Biomed Online, 7, pp.664~670 (2003)
  34. Wang Q, "Evaluation of oocyte quality: Morphological, cellular and molecular predictors", Reprod Fertil Dev, 19, pp.1~12 (2006)

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