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Başlık: Vitrification of pronuclear-stage mouse embryos on solid surface (SSV) versus in cryotube: Comparison of the effect of equilibration time and different sugars in the vitrification solution
Yazarlar: Bağış, Haydar
Mercan, Hande Odaman
Dinnyés, András
Uludağ Üniversitesi/Veteriner Fakültesi.
Sağırkaya, Hakan
AAH-8821-2021
6602400461
Anahtar kelimeler: Biochemistry & molecular biology
Cell biology
Developmental biology
Reproductive biology
Sugarcane streak virus
Mice
Pronuclear-stage embryos
Raffinose
Trehalose
Vitrification
In-vitro fertilization
Ethylene-glycol
Transgenic mice
Bovine oocytes
Developmental stage
Low toxicity
Rapid method
Cryopreservation
Survival
Yayın Tarihi: Şub-2004
Yayıncı: Wiley
Atıf: Bağış, H. vd. (2004). “Vitrification of pronuclear-stage mouse embryos on solid surface (SSV) versus in cryotube: Comparison of the effect of equilibration time and different sugars in the vitrification solution”. Molecular Reproduction and Development, 67(2), 186-192.
Özet: The cryopreservation of pronuclear-stage embryos has particular importance in transgenic technology and human assisted reproductive technology (ART). The objective of this study was to improve the efficiency of cryopreservation of pronuclear-stage mouse embryos. Two vitrification methods (solid surface vitrification (SSV) vs. vitrification in cryotube) have been compared with special emphasis on the effect of the exposure of the embryos to the solutions for various times and the sugar content (trehalose, sucrose, or raffinose) of the vitrification solutions. Pronuclear-stage embryos were either exposed to I M dimethyl sulfoxide (DMSO)+1 M propylene-glycol (PG) solution for 2, 5, 10, or 15 min or not exposed to this "equilibration" solution. The vitrification solutions consisted of 2.75 M DMSO and 2.75 M PG in M2 medium supplemented with 1 M trehalose (DPT), 1 M sucrose (DPS), or 1 M raffinose (DPR). In the cryotube method, groups of 15-25 embryos were transferred into a 1.8 ml cryotube containing 30 mul of DPT, DPS, or DPR. After 30 sec, the cryotubes were directly plunged into liquid nitrogen (LN2) and stored for 1 day to I month. Vitrified samples were warmed by immersing the cryotubes in a 40degreesC water bath and then immediately diluted with 300 mul of 0.3 M trehalose, sucrose, or raffinose in M2. In the SSV method, after equilibration 15-20 embryos were exposed to DPT, DPS, or DPR solutions for around 20 sec before being dropped in 2-mul drops onto a pre-cooled (-150 to -180degreesC) metal surface. Vitrified droplets were stored in cryovials in LN2. Warming was performed by transferring the vitrified droplets into 0.3 M solutions of trehalose, sucrose, or raffinose at 37degreesC, respectively. Results showed that both SSV and cryotube vitrification methods can result in high rates of in vitro blastocyst development (up to 58.3 and 68.5% with DPR, respectively), not statistically different from that of the controls (58.3 and 64.4%). Even without the equilibration step prior to vitrification, relatively high-survival rates have been achieved, except for the DPS solution. In conclusion, vitrification of pronuclear-stage mouse embryos can result in high rates of in vitro development to blastocyst, and the use of raffinose in the vitrification solution is advantageous to improve cryosurvival.
URI: https://doi.org/10.1002/mrd.10388
https://onlinelibrary.wiley.com/doi/10.1002/mrd.10388
http://hdl.handle.net/11452/21599
ISSN: 1040-452X
Koleksiyonlarda Görünür:Scopus
Web of Science

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