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

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 preimplantation genetic diagnosis for the genesis of animal models for animal disorders from embryos. In this study, female CB6 F1 (Balb/cXC57b1/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 degrees 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% O-2 and 90% air at 37 degrees 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.