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Journal of Animal and Veterinary Advances
Year: 2011  |  Volume: 10  |  Issue: 22  |  Page No.: 3027 - 3030

Optimal Method of Mouse Blastomere Biopsy: In vitro Developmental Potential of the Biopsied Embryo to Blastocyst Stage after Aspirated Eight-Cell Mouse Embryos

Ali Cihan Taskin, Tolga Akkoc, Arzu Tas Caputcu, Sezen Arat and Haydar Bagis    

Abstract: The researchers investigated the effect of blastocyst development and quality of blastomere aspiration techniques on eight-cell mouse embryos. The results clearly indicate that the in vitro development of biopsied mouse embryos depended on the suitable aspiration method. This method related to pre-implantation genetic diagnosis for the genesis of animal models for animal disorders from embryos. In this study, female CB6 F1(Balb/cXC57bl/j) hybrid mice were superovulated with hormones and superovulated females were sacrified approximately 68 h after hCG administration. About eight-cell embryos were recovered from oviducts of sacrified mouse in M2 medium. Before biopsy, all embryos were incubated to decrease cell to cell contacts to microdrops of Ca2+/Mg2 + free QAM HTF (3 mg mL-1 BSA Fraction V) with HEPES for 90 min at 37°C. A single blastomere of eight-cell embryos were aspirated by the aspiration pippetes under inverted microscope (4X). After biyopsy, embryos were cultured in SAGE medium supplemented with 5% CO2, 5% O2 and 90% air at 37°C for up to expanded blastocyst stage. After culture, 267 biopsied embryos out of 234 (88.32±7.5%) expanded blastocysts developed from the biopsy group and total cell number mean 67.8±17.2% were recorded in the experiment group and 126 non-biopsied embryos out of 118 (94.4±7.78%) expanded blastocysts developed from control embryos and total cell number mean 70±15.4% was recorded in the control group. In the present study, there was no difference in blastocysts developmental rates at post biopsy group and control group (p = 0.05). In conclusion, the biopsy the method described here is an optimal method of blastomere aspiration on in vivo mouse embryos.

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