Abstract
In vitro produced porcine embryos have potential application in reproductive biotechnology. However, their development potential has been very low. This study evaluated the in vitro developmental ability and quality of cloned and parthenogenetic porcine embryos having 2-4 cells or 5-8 cells on Day 2 of in vitro culture. Analysis of results showed that 2 to 4 cell embryos had higher ability to form blastocysts than 5 to 8 cell embryos (p<0.05). Blastocysts produced from culture of 2 to 4 cell embryos also contained higher cell numbers and had lower BAX:BCLxL transcript ratio than those produced from 5 to 8 cell embryos (p<0.05), thereby suggesting 2 to 4 cell embryos have higher development potential. Further investigation revealed that 5 to 8 cell embryos had higher incidence (100±0.0%) of blastomeric fragmentation than 2 to 4 cell embryos (15.2±5.5% for parthenogenetic and 27.7±7.1% for cloned embryos). This suggests that low development potential of 5 to 8 cell embryos was associated with blastomeric fragmentation. In conclusion, we have shown that morphological selection of embryos based on cell number on Day 2 of in vitro culture could offer a practical and valuable non-invasive means to select good quality porcine embryos.
Highlights
In vitro produced porcine embryos have potential application in reproductive biotechnology
Day 2 of oocytes were incubated for 20 min in TCM-199 medium in vitro culture (IVC) was chosen for analysis because embryo supplemented with 10% (v/v) fetal bovine serum (FBS) and 7.5 μg/ml transfer in the porcine is most commonly done at the 2 to 4 cytochalasin B (CB) and 5 μg/ml Hoechst 33342
At 168 h post activation, blastocysts were harvested for assessment of embryo quality by Hoechst 33342 staining for counting total cell number, differential staining for inner cell mass (ICM) rate, and real time quantitative reverse transcriptase polymerase chain reaction analysis for BAX:BCL-xL transcripts
Summary
Sci. 22(4):483491 cell number, ICM rate and fetal development (McKiernan Preparation of donor cells and Bavister 1994; Racowsky et al, 2000) It is Donor cells for nuclear transfer were prepared as plausible that counting the cell number of embryos on a described earlier (Uhm et al, 2000a). Have earlier shown that development characteristics of parthenogenetic embryos resembled those of IVF and Somatic cell nuclear transfer (SCNT). Day 2 of oocytes were incubated for 20 min in TCM-199 medium IVC was chosen for analysis because embryo supplemented with 10% (v/v) FBS and 7.5 μg/ml transfer in the porcine is most commonly done at the 2 to 4 cytochalasin B (CB) and 5 μg/ml Hoechst 33342. Ooplasm) using a beveled pipette (25 μm internal diameter) in HEPES buffered TCM medium supplemented with 10%
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
