| Objectives The inner cell mass were divided into three grades: A,B and C,which werein mouse parthenogenetic blastocyst(experimental group)and spermatozoa combined with blastocyst(control group).Through the research of the potential development ability of inner cell massto embryonic stem cells,to explore the characteristics of sexual development and development potential of parthenogenetic embryos and provide experimental data for the follow-up differentiation ability of parthenogenetic embryonic stem cells.Methods Female mice were used to promote ovulation by HMG and HCG combined method.MII eggs were activated by the formation of parthenogenetic embryos in the experimental group and fertilized by natural fertilization in the control group.Parthenogenetic embryos and fertilized eggs were cultured into blastocysts in vitro.Then,the cell clusters in the blastocysts were divided into three groups: A,B and C.,A,B,C three levels of cell clusters were respectively subcultured into stem cells.The activities of alkaline phosphatase and telomerase in the embryonic stem cell markers were detected and the ultrastructure of embryonic stem cells was observed by transmission electron microscopy.Results The expression level of alkaline phosphatase in the embryonic stem cells of A and B cells in the experimental group was the same as that in the control group(68.39 ± 7.92,65.79 ± 7.42 in the A group and 66.5 ± 8.32,66.16 ± 8.68 in the B group,P> 0.05).The expression level of alkaline phosphatase in embryonic stem cells of group C was significantly higher than that in group C(24.98 ± 4.37,44.26 ± 4.19 in group C),and the difference was Statistically significant(P <0.05).In the experimental group,The expression levels of alkaline phosphatase in the parthenogenetic embryonic stem cells in the A-level and B-class cell clusters were the same(Grade A 68.39 ± 7.92;Grade B 66.5 ± 8.32),the difference was not significant(P> 0.05).The expression level of alkaline phosphatase in the parthenogenetic embryonic stem cells in the C-class cell mass was lower than that of grade A,grade B(Grade C24.98±4.37),the difference was statistically significant(P <0.05).The expression level of telomerase in the embryonic stem cells of the experimental group was the same as that in the control group(A-level two groups of 10.18 ± 1.52,10.00 ± 1.24;B-level two groups 10.00 ± 1.50,10.11 ± 1.43),the difference was not significant(P> 0.05).The expression levels of telomerase in embryonic stem cell markers were significantly different between the two groups(5.67 ± 1.43,8.94 ± 0.99),the experimental group was lower,the difference was significant(P <0.05).In the experimental group,the level of telomerase expression of the parthenogenetic embryonic stem cell markers in the A-level and B-level cell clusters of the guinea pigs were the same(Grade A 10.18 ± 1.52,Grade B 10.00 ± 1.50),the difference was not significant(P> 0.05).The expression level of telomerase in the parthenogenetic embryonic stem cells formed by the C-levelcell clusters was lower than that of grade A and grade B(Grade C 5.67 ± 1.43),the difference was statistically significant(P <0.05).under transmission electron microscopy can be observed,in mouse parthenire blastocysts A,B grade inner cell mass that the ultrastructure of the parthenogenetic embryonic stem cells remained well,shape is simple and regular,cell gap is basically not visible,the cortex contains a large number of spherical mitochondria,volume is small and quantity,form is normal,the cytoplasm contains only a small amount of vacuoles;The stem cells ultrastructureformed from C-level cell clustersby parthenogenetic activation were irregular.The number of ribosomes is small and the morphology is relatively irregular,The volume was significantly swollen and rounded,the apoptosis was more common,the cell matrix increased shallow,the ridge decreased or disappeared,part of the cytoplasm of mitochondrial swelling lost normal form,part of the shape,the size of the vacuoles.Conclusions 1.The expression activity of parthenogenetic embryonic stem cells alkaline phosphatase,telomerase formed in mouse parthenire blastocysts A,B grade inner cell mass was the same.The expression activity of parthenogenetic embryonic stem cells alkaline phosphatase,telomerase formed in C grade inner cell mass lower than them formerd in A,B grade inner cell mass.2.Mouse parthenogenetic activated blastocyst has good development potential and subsequent differentiation ability,A,B grade inner cell mass have a good following development potential especially in the A grade. |
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