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. 2010 Feb 2;9(2):115-120.
doi: 10.1007/s12522-010-0045-6. eCollection 2010 Jun.

Vitrification of canine cumulus-oocyte complexes in DAP213 with a cryotop holder

Yasuyuki Abe  1 , Tomoyoshi Asano  1 , Mohammed Ali  1 , Hiroshi Suzuki  1
Affiliations

Vitrification of canine cumulus-oocyte complexes in DAP213 with a cryotop holder

Yasuyuki Abe et al. Reprod Med Biol. .

Abstract

Purpose: The effects of the cryoprotectant and the container (holder) used for the vitrification of canine germinal vesicle stage oocytes were examined to improve the cryopreservation method for canine oocytes and embryos.

Methods: Canine cumulus-oocyte complexes (COCs) were collected from ovaries, and were vitrified with E30S (30% ethylene glycol and 0.5 M sucrose) or DAP213 (2 M dimethyl sulfoxide, 1 M acetamide, and 3 M propylene glycol) solution held by a cryotube or cryotop sheets. After warming, the oocytes were stained with propidium iodide for the assessment of their plasma membrane integrity.

Results: In all the vitrification groups, more than 65% of the vitrified oocytes displayed a normal morphology (E30S-top, 65.6%; DAP-tube, 67.3%; DAP-top, 80.0%). However, when assessed by propidium iodide staining, the viability of oocytes in the DAP-top group (43.6%) was higher than that in the E30S-top group (21.3%, P < 0.05). Furthermore, the viability of the oocytes in the DAP-top group (43.6%) was higher than that in the DAP-tube group (4.1%, P < 0.05).

Conclusions: These results suggest that a combination of DAP213 as the cryoprotectant and a cryotop sheet as the holder improved viability after the vitrification of canine oocytes at the germinal vesicle stage.

Keywords: Cryopreservation; Cryotop; DAP213; Dog; Oocyte.

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Figures

Figure 1
Figure 1
Morphological appearance of canine oocytes after vitrification by the DAP‐tube method. a All oocytes show a morphologically normal appearance under light microscopy; b however, propidium iodide (PI) staining revealed that the oocytes stained red were damaged (arrows). Bar 200 μm
Figure 2
Figure 2
a Cooling and b warming rates in the cryotop and cryotube holders. The temperature in the holders was measured using a thermo recorder
See this image and copyright information in PMC

References

    1. Rodrigues BA, Rodrigues JL. Responses of canine oocytes to in vitro maturation and in vitro fertilization outcome. Theriogenology, 2006, 66, 1667–1672 - DOI - PubMed
    1. Songsasen N, Wildt DE. Oocyte biology and challenges in developing in vitro maturation systems in the domestic dog. Anim Reprod Sci, 2007, 98, 2–22 - DOI - PMC - PubMed
    1. Abe Y, Lee DS, Kim SK, Suzuki H. Vitrification of canine oocytes. J Mammal Ova Res, 2008, 25, 32–36 - DOI
    1. Nakagata N. Survival of mouse morulae and blastocysts derived from in vitro fertilization after ultra rapid freezing. Jikken Dobutsu, 1993, 42, 229–231 - PubMed
    1. Ishijima T, Kobayashi Y, Lee DS, Ueta YY, Matsui M, Lee JY, Suwa Y, Miyahara K, Suzuki H. Cryopreservation of canine ovaries by vitrification. J Reprod Dev, 2006, 52, 293–299 - DOI - PubMed