Preliminary Study on the Influencing Factors of Mouse Embryo Transfer

doi:10.3969/j.issn.1674-5817.2020.06.004

Abstract

Abstract: Objective To analyze the influences of mouse strain, embryo transfer method and transferred embryos' quantity on the results of embryo transfer, and to further optimize and improve the efficiency of embryo transfer. Methods The embryos from 277 gene modified strains were obtained by superovulation and in vitro fertilization (IVF), then the different numbers of embryos were transferred into the oviducts of pseudo-pregnant female mice through unilateral or bilateral embryo transplantation. The effects of strains with different IVF rates, number of transplanted embryos and transfer methods on the litter size and rate were analyzed and compared. Results When the strains with IVF rate of 0%-39%, the litter size (4.61±1.92) and the litter rate [(19.21±9.70)%were significantly lower than those in other groups (both P<0.01). The litter rate was significantly higher in 60%-69% IVF rate group than that in the 90%-100% IVF rate group (P<0.05). When the number of transferred embryos was fewer than 5, the recipient female mice got pregnant, but the litter size was less, with easy occurrence of the phenomenon of cannibalism. When the number of transferred embryos was more than 10, the litter size and rate of each group were both higher. The litter rate was (46.67±11.56)% when the number of transferred embryos was about 15, which was significantly higher than those of other groups (P<0.01), indicating that the embryo utilization rate was the highest. The average litter size was significantly the largest (9.67±1.53) with P<0.01 when the number of transferred embryos was around 25. The litter size (8.33±0.58) and the litter rate [(27.33±2.31)%were both decreased when the number of embryos transferred increased to 30, indicating that the bearing capacity of uterus almost reached the maximum. There was no significant difference in litter size or rate between unilateral and bilateral transplantation (P>0.05). Conclusion The average litter size and rate are lower when the IVF rate is lower than 40%. The embryo utilization rate is the highest, when the number of transferred embryos is about 15. The litter size is the largest when the number of transferred embryos is about 25, and the bearing capacity of uterus is almost the maximum. There is no significant difference in the litter size or rate between the unilateral and bilateral transplantation.

Key words: Embryo transfer, In vitro fertilization, Influence factor, Mice

CLC Number:

Q95-33

GUI Fei, YANG Weiwei, SUN Xiaopin, GU Meier. Preliminary Study on the Influencing Factors of Mouse Embryo Transfer[J]. Laboratory Animal and Comparative Medicine, 2020, 40(6): 483-488.

Figures/Tables 2

References

[1] Chen S, Zhang M, Li L, et al.Loss of methylation of H19-imprinted gene derived from assisted reproductive technologies can be mitigated by cleavage-stage embryo transfer in mice[J]. J Assist Reprod Genet, 2019, 36(11):2259-2269. doi: 10.1007/s10815-019-01575-x.
[2] 杜凯, 李文光, 吴德国, 等. 小鼠胚胎移植技术优化及对巨细胞病毒净化研究[J]. 实验动物科学, 2019, 36(5):31-34. doi: 10.3969/j.issn.1006-6179.2019.05.006.
[3] 牛博文, 陈丽香, 朱孟敏, 等. C57BL/6J 背景基因修饰小鼠精子冷冻保种及品系恢复方法[J]. 实验动物与比较医学, 2019, 39(2):146-150. doi: 10.3969/j.issn.1674-5817.2019.02.014.
[4] Schmidt PM, Hansen CT, Wildt DE.Viability of frozen-thawed mouse embryos is affected by genotype[J]. Biol Reprod, 1985, 32(3):507-514. doi: 10.1095/biolreprod32.3.507.
[5] Byers SL, Payson SJ, Taft RA.Performance of ten inbred mouse strains following assisted reproductive technologies (ARTs)[J]. Theriogenology, 2006, 65(9):1716-1726. doi: 10.1016/j.theriogenology.2005.09.016.
[6] Dorsch M, Wittur I, Garrels W.Success of embryo transfer in mice with freshly collected and cryopreserved two-cell embryos with different genetic backgrounds correlated with the number of transferred embryos: A 5-year retrospective analysis[J]. Lab Anim, 2019, 53(6):577-586. doi: 10.1177/0023677219832922.
[7] Guan M, Bogani D, Marschall S, et al.In vitro fertilization in mice using the MBCD-GSH protocol[J]. Curr Protoc Mouse Biol, 2014, 4(2):67-83. doi: 10.1002/9780470942390.mo140059.
[8] Takeo T, Sztein J, Nakagata N.The CARD method for mouse sperm cryopreservation and in vitro fertilization using frozen-thawed sperm[J]. Methods Mol Biol, 2019, 1874:243-256. doi: 10.1007/978-1-4939-8831-0_14.
[9] 邓欣, 白璐, 李森, 等. 一种小鼠卵巢分离及卵母细胞培养装置: CN201711455059.X[P].2017-12-28.
[10] Ramos RL, Van Dine SE, George E, et al.Molecular layer heterotopia of the cerebellar vermis in mutant and transgenic mouse models on a C57BL/6 background[J]. Brain Res Bull, 2013, 97:63-68. doi: 10.1016/j.brainresbull.2013.05.001.
[11] Song H, Lim H, Das SK, et al.Dysregulation of EGF family of growth factors and COX-2 in the uterus during the preattachment and attachment reactions of the blastocyst with the luminal epithelium correlates with implantation failure in LIF-deficient mice[J]. Mol Endocrinol, 2000, 14(8):1147-1161. doi: 10.1210 / mend.14.8.0498.
[12] Salamonsen LA, Dimitriadis E, Jones RL, et al.Complex regulation of decidualization: A role for cytokines and proteases-a review[J]. Placenta, 2003, 24(Suppl A):S76-85. doi: 10.1053/plac.2002.0928.
[13] Johnson LW, Moffatt RJ, Bartol FF, et al.Optimization of embryo transfer protocols for mice[J]. Theriogenology, 1996, 46:1267-1276. doi: 10.1016/S0093-691X(96)00298-1.
[14] Rülicke T, Haenggli A, Rappold K, et al.No transuterine migration of fertilised ova after unilateral embryo transfer in mice[J]. Reprod Fert Dev, 2006, 18(8):885-891. doi: 10.1071/rd06054.
[15] Mahabir E, Volland R, Landsberger A, et al.Reproductive performance after unilateral or bilateral oviduct transfer of 2-cell embryos in mice[J]. J Am Assoc Lab Anim Sci, 2018, 57(2):110-114.