کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2073801 1544793 2009 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Effect of different methods of cryopreservation on the cytoskeletal integrity of dromedary camel (Camelus dromedarius) embryos
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم کشاورزی و بیولوژیک علوم دامی و جانورشناسی
پیش نمایش صفحه اول مقاله
Effect of different methods of cryopreservation on the cytoskeletal integrity of dromedary camel (Camelus dromedarius) embryos
چکیده انگلیسی

This study examined the effect of different methods of cryopreservation on the cytoskeletal integrity of camel embryos. A total of 32 embryos were recovered on Days 6 and 7 after ovulation and measured before being frozen using either a conventional slow-cooling technique (n = 12: six Day 6 and six Day 7 embryos) or vitrification (n = 12: four Day 6 and eight Day 7). The remaining 8 ‘control’ embryos (four Day 6 and four Day 7) were not cryopreserved but instead incubated in holding medium for 30 min. After thawing, warming or incubation, the embryos were stained with 4,6-diamino-2-phenylindole dihydrochloride (DAPI) to identify dead cells. Subsequently, the embryos were fixed in 4% paraformaldehyde, permeabilized and labelled with Alexa Fluor 488-Phalloidin to enable assessment of cytoskeleton integrity.Vitrified–warmed embryos contained a significantly higher percentage of dead cells than either conventionally frozen embryos or controls (P < 0.05). Although the proportion of dead cells in conventionally frozen embryos tended to be higher than in controls, the difference was not significant (P ≥ 0.07). Whereas embryo size did not affect the number of dead cells in conventionally frozen embryos, vitrified–warmed embryos >300 μm in diameter had a significantly higher percentage of dead cells than embryos ≤300 μm (P = 0.01). Cytoskeleton integrity was also affected by both freezing method and embryo diameter. All 8 control embryos had a Grade I cytoskeleton, compared with only 2/24 (8.3%) frozen or vitrified embryos. Of the 8 slow-frozen or vitrified embryos with a Grade III cytoskeleton post-thaw, 7 had been vitrified and 6 were larger (Day 7) embryos. These results indicate that while both slow-freezing and vitrification of camel embryos lead to cytoskeleton disruption and cell death, embryo quality is better preserved by slow-freezing.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Animal Reproduction Science - Volume 113, Issues 1–4, July 2009, Pages 196–204
نویسندگان
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