Addition of macromolecules to PZM-3 culture medium on the development and hatching of in vitro porcine embryos.

摘要

Several experiments were conducted to develop enhanced cultural conditions for in vitro-produced porcine embryos. All embryos were produced by in vitro maturation (IVM) and fertilization (IVF) of immature oocytes from abattoir-derived ovaries. In experiment I, IVF embryos were divided into 4 groups and cultured in the following media: 0% bovine serum albumin (BSA); 3% BSA; 0.05% polyvinyl alcohol (PVA); and 0.5% polyvinylpyrrolidone (PVP) added to the basal fluid cultural medium, porcine zygote medium 3 (PZM-3). The rates of embryonic development were higher in the group where the PZM-3 media had been supplemented with 3% BSA than the other groups. Although insignificantly different, the percentages of blastocysts and hatched blastocysts were 6.9 and 25.0% in the 3% BSA group and 1.2-6.4 and 0-16.7% in the other groups, respectively. In experiment II, 10% fetal bovine serum (FBS) was added to PZM-3 on day 0 of culture, and the development rate of blastocysts per day of culture was observed from days 0 to 5. The development rate of blastocysts was higher at 15.6% on day 4 than on any other day, and was significantly higher than on day 0 or day one (p<0.05). The development rate of hatched blastocysts was 26.7% on day 4 and was higher than on any other day. In experiment III, IVF embryos were cultured with different fluid media: I - PZM-3 + 0.3% BSA (0-7 days); II - PZM-3 + 0.3% BSA-> (day 4) PZM-3 + 10% FBS; III - PZM-3 + 0.3% BSA-> PZM-3 + 0.3% BSA + 10% FBS (dasy 4); and IV - PZM-3 + 0.3% BSA + 10% FBS (days 0-7). The development rates of blastocysts and hatched blastocysts were 21.5 and 53.1% in group III, respectively, which were significantly higher than in group IV with respect to blastocyst development (5.2%, p<0.05) but not hatched blastocysts (14.3%). The total cell number (TCN) of blastocysts in group III was higher at 37.8+or-16.1 than the other groups at 16.8+or-4.4-30.1+or-10.9. However, this was insignificantly different. The results showed that PZM-3 containing 0.3% BSA and supplemented with FBS during the later stage of culture on day 4 resulted to better TCNs and increased rate of hatched blastocysts.