| To explore the effects on the development of mouse embryos between different culture mediums,growth factors and cultured gas phase, the ICR mice fertilized embryos (pronuclear embryos) were for the test materials. The aim of this study was to find a suitable culture medium composition and cultivation environment of ICR mouse embryo, and optimize the culture system of mouse embryos in vitro. At the same time, it was for reference as well as other mammalian embryos in vitro.There are four experiments, and all experiments were replicated for more than 5 times. Cleavage rate, blastocyst rate here were denoted with the average±standard error of that average, while the number of blastocyst cells denoted with the average±standard deviation. Results were analyzed by SAS statistical software.Specific results were as follows: (1) comparative study of the effects of the mKSOM, HTF, CZB three culture mediums on development of mouse pronuclear embryos. The results showed that the cleavage rate between the three groups were 74.1±5.6%, 64.3±7.3%, 71.9±6.1%, and there were no significant differences (P> 0.05). The blastocyst rate among three groups were 15.0±6.8%, 8.1±4.9%, 19.9±7.7%, while total cell number of blastocyst were 66.4±21.7, 68.0±7.3, 66.7±16.9, no significant differences among them (P> 0.05), but the mKSOM group got better efficiency. (2)study the effects of each ml mKSOM cotaining 1ng, 5ng, 10ng EGF or 10ng bFGF, 100ng bFGF on in vitro development of mouse pronuclear embryos. We found that cleavage rates among three groups containing EGF were 69.0±10.1%, 73.2±5.9%, 72.2±8.1%, the difference between these groups was not significant (P> 0.05). For the blastocyst rates, 26.6±12.9%, 18.2±7.1%, 29.2±15.5%, 10ng/ml EGF group was slightly higher than the other two groups (P> 0.05). Total cell number of blastocysts between three groups were 62.5±19.9, 51.0±9.7, 54.8±15.4, 1ng/mlEGF group was slightly higher than the other two groups, but the difference was not significant (P> 0.05). The cleavage rates of the two groups cotanning bFGF were 62.9±9.4%, 59.0±8.9%, without significant differences (P> 0.05). While blastocyst rate of the two groups were 23.6±7.8%, 25.4±7.3%, no significant differences (P> 0.05). (3) The effects of the micropore culture system (well of the well system, WOW) and group culture system on development of mouse prokaryotic embryos. The results revealed that the cleavage rate of the two groups were 93.6±2.4% and 83.2±4.8%, no significant difference (P> 0.05). The blastocyst rate of the two were 74.6±5.1% and 38.2±6.6%, and the total cell number of blastocyst were 76.2±2.7, 58.7±4.9. WOW system was significantly higher than the group culture, and there were significant differences (P <0.05). (4)The effects of Lung air (lung), 5% oxygen, 20% oxygen three different gas phase cultivation on development of mouse prokaryotic embryos. We found that cleavage rate of mKSOM group were 63.2±5.8%, 69.7±5.8%, 59.7±5.8%, There were no significant differences between the three gas phase cultivation (P> 0.05). Blastocyst rates of them were 6.7±5.1%, 22.6±5.1% and 6.1±5.1%, so 5% oxygen group was significantly higher than the other two groups (P <0.05). But difference between the other two groups was not significant. The total blastocyst cells respectively were 39.3±7.1, 7.2±5.8, 66.8±7.1, 20% oxygen group was significantly higher than the other two groups (P <0.05), while the difference between the other two groups was not significant. The HTF group's cleavage rates respectively were 44.1±5.8%, 60.2±5.8%, 54.8±5.8%, and each group had no not remarkable differences (P> 0.05). The blastocyst rate respectively were 0±0.0%, 6.7±5.1% and 2.8±5.1%, meanwhile the total cell number of blastocyst respectively were 0±0.0, 23.7±8.2, 19.0±8.2. Differences between the groups were not remarkable (P> 0.05). The cleavage rates in CZB group were 64.4±5.8%, 59.7±5.8%, 57.9±6.2%, Differences were not significant (P> 0.05). The blastocyst rates were 16.2±5.1%, 28.7±5.1% and 16.4±5.4%, and the differences between the groups was not significant (P> 0.05). But 5% oxygen culture group had the highest blastocyst rate. The total cell number of blastocyst were 53.8±3.3%, 43.3±3.6%, 49.7±5.3%. 20% oxygen group had no significant differences between the other two groups, however difference between the other two groups was significant (P <0.05). In addition, under the conditions of three kinds of gas in CZB medium it showed higher blastocyst rates, although the differences were not statistically significant (P> 0.05). However, the total cell number of blastocyst significantly increased under the lung and 20% O2 gas ( 53.8±3.3%, 49.7±5.3% vs 43.3±3.6%, P <0.05). That indicated 5% oxygen group and the group of lung gas can be used to cultivate the mouse pronuclear embryos in vitro.In conclusion, it would be more efficient by the choice of (1) mKSOM medium with the use of WOW with 5% in oxygen under the conditions of the gas, or (2) CZB + WOW + Lung air in the development of the mouse pronuclear embryos in vitro.
|
PDF Full Text Request