共培养对小鼠孤雌生殖胚胎体外发育的影响

Yanyan Qiao, Chunxiao Zhang, Yuanyuan Jiang
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摘要

目的研究输卵管上皮细胞体外共培养对孤雌胚胎发育的影响。方法胰酶收集兔输卵管上皮细胞,37℃,5%CO2培养。6 ~ 8周龄昆明白种雌性小鼠控制卵巢过度刺激。用7%乙醇和5 mg/L细胞松弛素b激活成熟卵母细胞孤雌生殖,对照组(单纯培养)和实验组(兔输卵管上皮细胞P0和P1、P2、P3、P4共培养体系)分别培养孤雌生殖激活卵。1 ~ 5 d后,观察小鼠孤雌胚胎发育为2细胞、4细胞、8细胞和囊胚。结果与对照组相比,小鼠孤雌活化卵与原代P0和传代P1、P2、P3兔输卵管上皮细胞共培养,2细胞和4细胞的形成率差异无统计学意义(P < 0.05)。8细胞形成率显著提高(57.5%比80.2%、85.0%、81.8%比76.8%,χ2=14.389、23.803、17.319、10.078,P<0.01)。囊胚形成率显著提高(34.5%比57.9%、62.9%、58.3%、50.4%,χ2=13.126、20.094、13.857、6.116,P<0.05)。结论输卵管上皮细胞体外共培养可明显提高孤雌胚胎的发育潜能。关键词:孤雌胚胎;共培养;囊胚率
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Effects of co-culture on in vitro development of parthenogenetic embryos in mice
Objective To study the effect of co-culture of tubal epithelial cells on the development of parthenogenetic embryos in vitro. Methods Rabbit oviduct epithelial cells were collected by trypsin and cultured at 37 ℃ and 5%CO2. Six to eight weeks age of kunming white female mice were controlled ovarian hyperstimulation. Mature oocytes were parthenogenetic activated by 7% ethanol and 5 mg/L cytochalasin B. Parthenogenetic activated eggs were cultured in the control group (simple culture), and the experimental group (rabbit oviduct epithelial cells P0 and P1, P2, P3, P4 co-culture system). After 1-5 d, the mice parthenogenetic embryos to 2-cell, 4-cell, 8-cell and the blastocyst were observed. Results Compared with the control group, the parthenogenetic activated eggs of mice were co-cultured with primary P0 and pass-through P1, P2 and P3 rabbit oviduct epithelial cells, and there was no significant difference in the formation rate of 2-cell and 4-cell (P>0.05). The 8-cell formation rate was significantly increased (57.5% vs. 80.2%, 85.0%, 81.8% vs. 76.8%, χ2=14.389, 23.803, 17.319, 10.078, P<0.01). Blastocyst formation rate increased significantly (34.5% vs. 57.9%, 62.9%, 58.3%, 50.4%, χ2=13.126, 20.094, 13.857, 6.116, P<0.05). Conclusion Co-culture of oviduct epithelial cells can obviously improve the development potential of parthenogenetic embryos in vitro. Key words: Parthenogenetic embryos; Co-culture; Blastocyst rate
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