玻璃化冷冻胚胎出生小鼠卵巢发育情况及卵巢组织中生长分化因子9的表达

Study on ovary development in vitrification of embryos born mice and expression of growth differentiation factor 9

目的探讨玻璃化冷冻胚胎出生小鼠的卵巢发育情况及卵巢组织中生长分化因子9（GDF-9）的表达。方法将玻璃化冷冻复苏胚胎（玻璃化冷冻组）和新鲜胚胎（新鲜胚胎组）分别移植入假孕小鼠子宫内，两组出生后的雌性子鼠分别于第3、7、14、21、28、60天处死，取卵巢组织，每组每个时间点各6只子鼠。对卵巢组织采用HE染色观察卵巢发育情况，采用荧光定量逆转录（RT）PCR技术和蛋白印迹法检测卵巢组织中生长分化因子9（GDF-9）mRNA和蛋白的表达水平。结果HE染色结果显示，两组子鼠卵巢发育无明显异常。子鼠出生后第3、7、14、21、28、60天，玻璃化冷冻组子鼠卵巢组织中GDF-9mRNA表达水平分别为：0．14±0．07、0．42±0．16、1．00±0．24、1．59±0．28、2．05±0．32、2．23±0．21，新鲜胚胎组分别为：0．13±0．06、0．45±0．18、1．00±0．21、1．56±0．26、2．01±0．37、2．26±0．23，两组分别比较，差异均无统计学意义（P〉0．05）；玻璃化冷冻组各时间点子鼠卵巢组织中GDF-9蛋白表达水平分别为：0．040±0．030、0．120±0．060、0．170±0．030、0．250±0．040、0．320±0．060、0．330±0．010，新鲜胚胎组分别为：0．030±0．020、0．110±0．040、0．150±0．010、0．210±0．020、0．360±0．070、0．350±0．030，两组分别比较，差异均无统计学意义（P〉0．05）。结论玻璃化冷冻胚胎出生小鼠的卵巢形态及GDF一9表达无明显变化。

Objective To study ovarian development in vitrification of embryos born mice and expression of growth differentiation factor 9 （ GDF-9 ） in its. Methods The vitrification recovery embryos （vitrified-embryo group ） and fresh embryos （fresh-embryo group ） were transplanted into pseudopregnant mice, respectively. The female offspring mice in two groups were sacrificed on the 3rd, 7th, 14th, 21th, 28th and 60th day after birth, the ovarian tissues were taken, 6 mice in each time point of each group. The ovarian development was observed by HE staining, the expression of GDF-9 mRNA and protein at each time point of two groups were detected by reverse transcription（RT）-PCR and western blot. Results HE staining showed that no abnormal ovarian development was observed in offsprings at two groups. On the 3rd, 7th, 14th, 21st, 28th and 60th day after birth, the expression of GDF-9 mRNA in vitrified-embryo group were 0. 14 ± 0. 07, 0. 42 ± 0. 16, 1.00 ± 0. 24, 1.59 ± 0. 28, 2.05± 0. 32 and 2. 23 ± 0. 21, respectively, which in freshembryo group were 0. 13 ±0. 06, 0. 45 ±0. 18, 1.00 ±0. 21, 1.56 ±0. 26, 2.01±0. 37 and 2. 26 ±0. 23, respectively, there was no statistical difference between two groups （ P 〉 0. 05 ） ; the expression of GDF-9 protein in vitrified-embryo group were 0. 040± 0. 030, 0. 120 ± 0. 060, 0. 170 ± 0. 030, 0. 250 ± 0. 040, 0. 320 ± 0. 060 and 0. 330 ± 0. 010, respectively, which in fresh-embryo group were 0. 030 ± 0. 020, 0. 1 10 ± 0. 040, 0. 150 ± 0. 010, 0. 210 ± 0. 020, 0. 360 ± 0. 070 and 0. 350 ± 0. 030, respectively, there was no statistical difference between two groups （ P 〉 0. 05 ）. Conclusion The ovarian morphology in vitrification of embryos born mice and expression of GDF-9 in ovary has no any obvious change.