Ploidy of Bovine Nuclear Transfer Blastocysts Blastomere Donors

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

The higher rate of embryonic loss in nuclear transfer compared to in vitro produced embryos may be due to chromosome abnormalities that occur during preimplantation in vitro devel- opment. Because little is known about ploidy errors in nuclear transfer embryos, this was ex- amined using embryos reconstructed from in vitro produced embryo donors. In vitro matured oocytes were enucleated and then activated using calcium ionophore A23187 followed by 6- dimethylaminopurine (6-DMAP). Subsequently, embryos were reconstructed using blas- tomeres from day 4–5 in vitro produced donors. The embryos were cultured until day 7 at which time blastocyst nuclei were extracted and chromosome abnormalities were evaluated by fluorescent in situ hybridization using two probes that bind to the subcentromeric regions on chromosomes 6 and 7. In 16 nuclear transfer blastocysts generated from 5 donor embryos, 53.8 6 20.2 (mean SD) nuclei/embryo were examined. Of these 16, 7 embryos (43.8 were potentially abnormal because in these, 1.1 1.4 5.3 7.5 26.3 30.4 and 66.2% % of the nuclei had a chromosome composition deviating from the diploid condition, indicating a wide degree of variation between embryos. These errors comprised mainly triploid (8.2 6 10.3 [0–26.3]: SD [range]) and tetraploid (10.6 6 19.9 [0–54.9]) nuclei with other ploidy com- binations accounting for only 0.9 6 2.1 [0–2.1]% of deviant nuclei. The proportion of com- pletely normal nuclear transfer embryos was no less than those produced by in vitro fertil- ization but the distribution of chromosome abnormalities was different (p 5 0.0002). In conclusion, nuclear transfer embryos reconstructed using blastomere cells can produce over 50% blastocysts with a diploid chromosome complement. However, the contribution of chro- mosome abnormalities to embryonic loss in the remaining embryos deserves further investi- gation
OriginalsprogUdefineret/Ukendt
TidsskriftCloning
Vol/bind2
Udgave nummer2
Sider (fra-til)63-69
Antal sider7
StatusUdgivet - 2000

ID: 141568142