Open AccessVitrification of canine cumulus–oocyte complexes in DAP213 with a cryotop holder
Author(s) -
Abe Yasuyuki,
Asano Tomoyoshi,
Ali Mohammed,
Suzuki Hiroshi
Publication year - 2010
Publication title -
reproductive medicine and biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.005
H-Index - 22
eISSN - 1447-0578
pISSN - 1445-5781
DOI - 10.1007/s12522-010-0045-6
Subject(s) - germinal vesicle , cryoprotectant , vitrification , propidium iodide , andrology , cryopreservation , chemistry , oocyte , dimethyl sulfoxide , embryo , biology , biochemistry , medicine , microbiology and biotechnology , apoptosis , organic chemistry , programmed cell death
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.