Browse > Article
http://dx.doi.org/10.12717/DR.2018.22.3.245

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

Hwang, In-Sul (Animal Biotechnology Division, National Institute of Animal Science)
Park, Mi-Ryung (Animal Biotechnology Division, National Institute of Animal Science)
Kwak, Tae-Uk (Animal Biotechnology Division, National Institute of Animal Science)
Park, Sang-Hyun (Animal Biotechnology Division, National Institute of Animal Science)
Lim, Ji-Hyun (Animal Biotechnology Division, National Institute of Animal Science)
Kim, Sung Woo (Animal Genetics Resources Research Center, National Institute of Animal Science)
Hwang, Seongsoo (Animal Biotechnology Division, National Institute of Animal Science)
Publication Information
Development and Reproduction / v.22, no.3, 2018 , pp. 245-252 More about this Journal
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 cytochalasin 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 postwarming pig oocytes were analyzed by fluorescein diacetate (FDA) assays with 4 classifications. In results, post-warming oocytes 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%), significantly (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 refresh the post-warming pig oocyte resulting its improved survival rates.
Keywords
Pig oocyte; Vitrification; Cytochalasin B; Cryoinjury; Viability;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Balaban B, Barut T, Urman B (2012) Assessment of oocyte quality. In: Nagy ZP, Varghese AC, Agarwal A (eds.), Practical Manual of In Vitro Fertilization: Advanced Methods and Novel Devices, Springer, New York.
2 Balaban B, Urman B, Sertac A, Alatas C, Aksoy S, Mercan R (1998) Oocyte morphology does not affect fertilization rate, embryo quality and implantation rate after intracytoplasmic sperm injection. Hum Reprod 13:3431-3433.   DOI
3 Boyer P, Tourame P, Gervoise-Boyer M (2012) Oocyte vitrification and its impact on the clinical practice of assisted reproduction. Gynecol Obstet Fertil 40:687-690.   DOI
4 Chasombat J, Nagai T, Parnpai R, Vongpralub T (2015) Pretreatment of in vitro matured bovine oocytes with docetaxel before vitrification: Effects on cytoskeleton integrity and developmental ability after warming. Cryobiology 71:216-223.   DOI
5 Coticchio G, Bromfield JJ, Sciajno R, Gambardella A, Scaravelli G, Borini A, Albertini DF (2009) Vitrification may increase the rate of chromosome misalignment in the metaphase II spindle of human mature oocytes. Reprod Biomed Online 3:29-34.
6 Dai J, Wu C, Muneri CW, Niu Y, Zhang S, Rui R, Zhang D (2015) Changes in mitochondrial function in porcine vitrified MII-stage oocytes and their impacts on apoptosis and developmental ability. Cryobiology 41:291-298.
7 De Sutter P, Dozortsev D, Qian C, Dhont M (1996) Oocyte morphology does not correlate with fertilization rate and embryo quality after intracytoplasmic sperm injection. Hum Reprod 11:595-597.   DOI
8 dela Pena EC, Takahashi Y, Atabay EC, Katagiri S, Nagano M (2001) Vitrification of mouse oocytes in ethylene glycol-raffinose solution: Effects of preexposure to ethylene glycol or raffinose on oocyte viability. Cryobiology 42:103-111.   DOI
9 Didion BA, Pomp D, Martin MJ, Homanics GE, Markert CL (1990) Observations on the cooling and cryopreservation of pig oocytes at the germinal vesicle stage. J Anim Sci 68:2803-2810.   DOI
10 Fabbri R, Porcu E, Marsella T, Primavera MR, Seracchioli R, Ciotti PM, Magrini O, Venturoli S, Flamigni C (1998) Oocyte cryopreservation. Hum Reprod 13:98-108.
11 Fuku E, Xia L, Downey BR (1995) Ultrastructural changes in bovine oocytes cryopreserved by vitrification. Cryobiology 32:139-156.   DOI
12 Hamamah S (2005) Oocyte and embryo quality: Is their morphology a good criterion? J Gynecol Obstet Biol Reprod (Paris) 34:5S38-5S41.
13 Hara H, Hwang IS, Kagawa N, Kuwayama M, Hirabayashi M, Hochi S (2012) High incidence of multiple aster formation in vitrified-warmed bovine oocytes after in vitro fertilization. Theriogenology 77:908-915.   DOI
14 Hwang IS, Hara H, Chung HJ, Hirabayashi M, Hochi S (2013) Rescue of vitrified-warmed bovine oocytes with rho-associated coiled-coil kinase inhibitor. Biol Reprod 89:1-6.
15 Isachenko V, Soler C, Isachenko E, Perez-Sanchez F, Grishchenko V (1998) Vitrification of immature porcine oocytes: Effects of lipid droplets, temperature, cytoskeleton, and addition and removal of cryoprotectant. Cryobiology 36:250-253.   DOI
16 Hwang IS, Hochi S (2014) Recent progress in cryopreservation of bovine oocytes. Biomed Res Int 2014:570647.
17 Hwang IS, Kwon DJ, Kwak TU, Lee JW, Im GS, Hwang S (2016) Improved survival and developmental rates in vitrified-warmed pig oocytes after recovery culture with coenzyme Q10. Cryo Letters 37:59-67.
18 Hwang IS, Kwon DJ, Oh KB, Ock SA, Chung HJ, Cho IC, Lee JW, Im GS, Hwang S (2015) Production of cloned Korean native pig by somatic cell nuclear transfer. Dev Reprod 19:79-84.   DOI
19 Kikuchi K, Ekwall H, Tienthai P, Kawai Y, Noguchi J, Kaneko H, Rodriguez-Martinez H (2002) Morphological features of lipid droplet transition during porcine oocyte fertilisation and early embryonic development to blastocyst in vivo and in vitro. Zygote 10:355-366.   DOI
20 Kono T, Kwon OY, Nakahara T (1991) Development of vitrified mouse oocytes after in vitro fertilization. Cryobiology 28:50-54.   DOI
21 Lowther KM, Weitzman VN, Maier D, Mehlmann LM (2009) Maturation, fertilization, and the structure and function of the endoplasmic reticulum in cryopreserved mouse oocytes. Biol Reprod 81:147-154.
22 Marco-Jimenez F, Casares-Crespo L, Vincente JS (2012) Effect if cytochalasin B pre-treatment of in vitro matured porcine oocytes before vitrification. Cryo Letters 33:24-30.
23 Mohr LR, Trounson AO (1980) The use of fluorescein diacetate to assess embryo viability in the mouse. J Reprod Fertil 58:189-196.   DOI
24 Rojas C, Palomo MJ, Albarracin JL, Mogas T (2004) Vitrification of immature and in vitro matured pig oocytes: Study of distribution of chromosomes, microtubules, and actin microfilaments. Cryobiology 49:211-220.   DOI
25 Mullen SF, Fahy GM (2012) A chronologic review of mature oocyte vitrification research in cattle, pigs, and sheep. Theriogenology 78:1709-1719.   DOI
26 Noto V, Campo R, Roziers P, Gordts S (1991) Fluorescein diacetate assessment of embryo viability after ultrarapid freezing of human multipronucleate embryos. Fertil Steril 55:1171-1175.   DOI
27 Pitchayapipatkul J, Somfai T, Matoba S, Parnpai R, Nagai T, Geshi M, Vongpralub T (2017) Microtubule stabilisers docetaxel and paclitaxel reduce spindle damage and maintain the developmental competence of in vitro-mature bovine oocytes during vitrification. Reprod Fertil Dev 29:2028-2039.   DOI
28 Somfai T, Kikuchi K, Nagai T (2012) Factors affecting cryopreservation of porcine oocytes. J Reprod Dev 58:17-24.   DOI
29 Vallorani C, Spinaci M, Bucci D, Porcu E, Tamanini C, Galeati G (2012) Pig oocyte vitrification by Cryotop method and the activation of the apoptotic cascade. Anim Reprod Sci 135:68-74.   DOI
30 Van Soom A, Mateusen B, Leroy J, de Kruif A (2003) Assessment of mammalian embryo quality: What can we learn from embryo morphology? Reprod Biomed Online 7:664-670.   DOI
31 Wang Q, Sun Q-Y (2006) Evaluation of oocyte quality: Morphological, cellular and molecular predictors. Reprod Fertil Dev 19:1-12.
32 Amoushahi M, Salehnia M, Mowla SJ (2017) Vitrification of mouse MII oocyte decreases the mitochondrial DNA copy number, TFAM gene expression and mitochondrial enzyme activity. J Reprod Infertil 18:343-351.
33 Albarracin JL, Morato R, Rojas C, Mogas T (2005) Effects of vitrification in open pulled straws on the cytology of in vitro matured prepubertal and adult bovine oocytes. Theriogenology 63:890-901.   DOI
34 Almasi Turk S, Roozbehi A (2013) 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 16:599-601.