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http://dx.doi.org/10.12749/RDB.2015.39.3.77

Toxic Effects of Ethylene Glycol on Mammalian Embryo Survivability  

Kim, Hyun (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA)
Yu, Dae Jung (Livestock Research Institute, Jeollanam-do Agricultural Research & Extension Service)
Choe, Changyong (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA)
Seong, Hwan-Hoo (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA)
Publication Information
Reproductive and Developmental Biology / v.39, no.3, 2015 , pp. 77-81 More about this Journal
Abstract
This study was carried out to evaluate the effects of embryonic stage and toxicities of cryoprotectant on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology. Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and ethylene glycol (EG) were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or EG. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO ($68.1{\pm}24.1$) at the 2-cell stage was significantly lower than that were treated with EG ($81.2{\pm}27.0$) or the control ($99.0{\pm}18.3$) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the EG treated group (p<0.05). Both DMSO ($15.4{\pm}1.5$) and EG ($10.2{\pm}1.4$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.1{\pm}0.9$, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the EG treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or EG at room temperature. When comparing two cryoprotective agents, EG appeared to be less toxic than DMSO at least in a murine embryo model.
Keywords
Cryoprotectant; DMSO; EG; Toxicity; Embryo;
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1 Gilmore JA, McGann LE, Liu J, Gao DY, Peter AT, Kleinhans FW (1995) Effect of cryoprotectant solutes on water permeability of human spermatozoa. Biol Reprod 53:985-995.   DOI
2 Hey JM, MarFarlane DR (1996) Crystallization of ice in aqueous solutions of glycerol and dimethly sulphoxide. Cryobiology 33:205-216.   DOI
3 Kuleshova LL, Shaw JM (2001) Studies on replacing most of the penetrating cryoprotectant by polymers for embryo cryopreservation. Cryobiology 43:21-31.   DOI
4 Lovelock JE, Bishop MW (1959) Prevention of freezing damage to living cells by dimethyl sulphoxide. Nature 183:1394-1395.   DOI
5 Kim MK, Lee SJ, Uhm EU, Yoon SH, Park SP, Chung KS, Lim JH (1996) Cryopreservation of mouse IVF zygotes by vitrification. J Animal Reprod 20:119-126.
6 Lovelock JE, Bishop MW (1959) Prevention of freezing damage to living cells by dimethyl sulphoxide. Nature 183:1394-1395.   DOI
7 Mandelbaum J, Junca AM, Plachot M, Alnot MO, Tibi C, Cohen J, Salat-Baroux J (1988) Solutions provided by the freezing of embryos and questions posed by the freezing of human oocytes. Rev Fr Gynecol Obstet 83:619-622.
8 Martino A, Songsasen N, Leibo SP (1996) Development into blastocysts of bovine oocytes cryopreserved by ultrarapid cooling. Biol Reprod 54:1059-1069.   DOI
9 Mahadevan MM, Miller MM (1997) Deleterious effect of equilibration temperature on the toxicity of propanediol during cryopreservation of mouse zygotes. J Assist Reprod Genet 14:51-54.   DOI
10 Newton H, Fisher J, Arnold JR, Pegg DE, Faddy MJ, Gosden RG (1998) Permeation of human ovarian tissue with cryoprotective agents in preparation for cryopreservation. Hum Reprod 13:376-380.   DOI
11 Newton H, Pegg DE, Barrass R, Gosden RG (1999) Osmotically inactive volume, hydraulic conductivity, and permeability to dimethyl sulphoxide of human mature oocyte. J Reprod Fertil 117:27-33.   DOI
12 Nowshari MA, Nayudu PL, Hodges JK (1995) Effect of cryoprotectants and their concentration on postthaw survival and development of rapid frozen-thawed pronuclear stage mouse embryos. Hum Reprod 10:3237-3242.   DOI
13 Rayos AA, Takahashi Y, Hishinuma M, Kanagawa H (1994) Quick freezing of unfertilized mouse oocytes using ethylene glycol with sucrose or trehalose. J Reprod Fertil 24:100-123.
14 Schiewe MC, Hazeleger NL, Scliment DF, Maceda JP (1995) Physiological characterization of blastocyst hatching mechanisms by use of a mouse antihatching model. Fertil Steril 63:288-294.   DOI
15 Sutton RL (1991) Critical cooling rate to avoid ice crystallization in solutions of cryoprotective agents. J Chem Soc Faraday Trans 87:101-105.   DOI
16 Takagi M, Boediono A, Saha S, Suzuki T (1993) Survival rate of frozen-thawed bovine IVF embryos in relation to exposure time using various cryoprotectants. Cryobiology 30:306-312.   DOI
17 Yoon TK, Lee DR, Cha SK, Chung HM, Lee WS, Cha KY (2007) Survival rate of human oocytes and pregnancy outcome after vitrification using slush nitrogen in assisted reproductive technologies. Fertil Steril 88:952-956.   DOI
18 Candy CJ, Wood MJ, Whittingham DG (1997) Effect of cryoprotectants on the survival of follicles in frozen mouse ovaries. J Reprod Fertil 110:11-19.   DOI
19 Zhu SE, Kasai H, Otoge T, Sakurai T, Machida T (1993) Cryopreservation of expanded mouse blastocysts by vitrification in ethylene glycol-based solutions. J Reprod Fert 98:139-145.   DOI
20 Boutron P (1984) A more accurate determination of the quantity of ice crystallized at low cooling rates in the glycerol and 1,2-propandiol aqueous solutions: Comparison with equilibrium. Cryobiology 21:183-191.   DOI
21 Cohen J, Simons RE, Edwards RG, Fehilly CB, Fisher FB (1985) Pregnancies following the frozen storage of expanding human blastocysts. J In Vitro Fert Embryo Transf 2:59-64.   DOI
22 Demici B, Lomage J, Salle B, Frappart L, Franck M, Guerin JF (2001) Follicular viability and morphology of sheep ovaries after exposure to cryoprotectant and cryopreservation with different freezing protocols. Fertil Steril 75:754-762.   DOI
23 Dumoulin JC, Bergers-Janssen JM, Pieters MH, Enginsu ME, Geraedts JP, Evers JL (1994) The protective effects of polymers in the cryopreservation of human and mouse zonae pellucidae and embryos. Fertil Steril 62:793-798.   DOI
24 Friedler S, Giudice LC, Lamb EJ (1988) Cryopreservation of embryos and ova. Fertil Steril 49:743-764.   DOI
25 Gardner DK, Lane M (2000) Embryo culture systems. In "Handbook of In Vitro Fertilization" 2nd Ed CRC Press Boca Raton FL 2:205-264.