小鼠胚胎移植技术的优化及应用

doi:10.12300/j.issn.1674-5817.2026.069

实验动物与比较医学 ›› 2026, Vol. 46 ›› Issue (5): 750-758.DOI: 10.12300/j.issn.1674-5817.2026.069

小鼠胚胎移植技术的优化及应用

张娇玲(), 林阿萍, 许诗荧, 潘德全, 蔡毅君()()

^1. 翔安创新实验室, 传染病疫苗研发全国重点实验室, 厦门 361102
^2. 厦门大学公共卫生学院, 厦门 361102

收稿日期:2026-04-17 修回日期:2026-08-05 出版日期:2026-10-25 发布日期:2026-10-16
通讯作者: 蔡毅君（1983—），女，硕士，高级工程师，研究方向：基因编辑、动物模型构建。E-mail： yjcai@xmu.edu.cn。ORCID： 0009-0006-8086-9073
作者简介:
张娇玲（1997—），女，本科，技术员，研究方向：实验动物模型构建、生物净化和繁殖与保种研究。E-mail： jiaolingzhang@xmu.edu.cn。ORCID： 0009-0001-8096-1420
基金资助:
2024年度厦门市自然科学基金面上项目“基于hCAR转基因小鼠的病毒性胰腺炎模型研究及抗病毒药物评估”(3502Z202473106)

Optimization and Application of Mouse Embryo Transfer Technology

ZHANG Jiaoling(), LIN Aping, XU Shiying, PAN Dequan, CAI Yijun()()

^1. Xiang'an Biomedicine Laboratory, State Key Laboratory of Vaccines for Infectious Diseases, Xiamen 361102, China
^2. School of Public Health Xiamen University, Xiamen 361102, China

Received:2026-04-17 Revised:2026-08-05 Published:2026-10-25 Online:2026-10-16
Correspondence to:
CAI Yijun (ORCID: 0009-0006-8086-9073), E-mail: yjcai@xmu.edu.cn

摘要/Abstract

摘要：

目的优化小鼠胚胎移植的技术参数（包括输卵管开口方式、假孕受体体重、单只受体移植胚胎数量、单侧/双侧输卵管移植4项技术参数），提高胚胎移植效率（以胚胎移植后受体母鼠妊娠率作为评判指标），提高优质胚胎种质资源利用率（以胚胎移植后小鼠出生率作为评判指标）。方法以560只SPF级ICR小鼠和440只C57BL/6J小鼠为实验对象，通过体外受精获得2细胞胚胎，采用单因素控制变量法，系统探究输卵管开口方式、假孕受体体重、单只受体移植胚胎数量、单侧/双侧输卵管移植4种因素对小鼠胚胎移植妊娠率和出生率的影响。结果采用26G注射器针头打孔的输卵管开口方式，受体的妊娠率可达90%，显著高于维纳斯剪剪切开口组的43%（P<0.05）。体重29～35 g的假孕受体移植效果最优（受体妊娠率为90%、幼鼠出生率为44%，且无幼崽被母鼠压死的现象）。单只受体移植胚胎数量在16～25枚组的出生率达42%，为各组最高；26～35枚组的妊娠率达90%，且胚胎利用率达到最大；而胚胎数超过35枚后移植效率显著下降。单只受体的胚胎移植数量在24～26枚，可兼顾较高的妊娠率和出生率。小鼠胚胎经输卵管双侧移植的妊娠率和出生率较单侧移植略高，但差异无统计学意义（P>0.05）。结论选择体重29～35 g的假孕受体母鼠，采用26G注射器针头打孔的输卵管开口方式，单只受体移植胚胎的数量控制在24～26枚，能够有效提高胚胎移植成功率，实现优质胚胎种质资源的高效利用。

关键词: 胚胎移植, 体外受精, 技术优化, 移植效率, ICR小鼠, C57BL/6J小鼠

Abstract:

Objective To optimize the technical parameters of mouse embryo transfer (including oviduct opening method, pseudopregnant recipient body weight, number of embryos transferred per recipient, and unilateral vs. bilateral oviduct transfer), improve the efficiency of embryo transfer (the pregnancy rate of recipient female mice after embryo transfer is used as the evaluation index), and improve the utilization rate of high-quality embryo germplasm resources (the birth rate of mice after embryo transfer is used as the evaluation index). Methods 560 SPF-grade ICR mice and 440 C57BL/6J mice were used as experimental animals. 2-cell embryos were obtained through in vitro fertilization. A single-factor controlled variable method was used to systematically investigate the effects of four factors, including oviduct opening method, pseudopregnant recipient body weight, number of embryos transferred per recipient, and unilateral vs. bilateral oviduct transfer, on the pregnancy rate and birth rate of mouse embryo transfer. Results The pregnancy rate of recipients with an opening made in the oviduct using a 26G syringe needle reached 90%, which was significantly higher than the 43% in the group with oviduct openings cut by Vannas scissors (P<0.05). Pseudopregnant recipients weighing 29-35 g exhibited optimal embryo transfer outcomes, with a pregnancy rate of 90% and a birth rate of 44%, and no pups were crushed by the recipient mothers. The highest birth rate of 42% was observed in the group receiving 16-25 embryos per recipient. Transferring 26-35 embryos per recipient achieved a 90% pregnancy rate and maximized embryo utilization efficiency. However, transfer efficiency decreased significantly when more than 35 embryos were transferred. Transfer of 24-26 embryos per recipient balanced relatively high pregnancy and birth rates. Bilateral oviduct transfer of mouse embryos yielded slightly higher pregnancy and birth rates than unilateral transfer, but the difference was not statistically significant (P>0.05). Conclusion Selecting pseudopregnant recipient female mice weighing 29-35 g, using a 26G syringe needle to make an opening in the oviduct, and controlling the number of transferred embryos at 24-26 per recipient can effectively improve the success rate of embryo transfer and achieve efficient utilization of high-quality embryo germplasm resources.

Key words: Embryo transfer, In vitro fertilization, Technical optimization, Transfer efficiency, ICR mice, C57BL/6J mice

中图分类号:

张娇玲,林阿萍,许诗荧,等. 小鼠胚胎移植技术的优化及应用[J]. 实验动物与比较医学, 2026, 46(5): 750-758. DOI: 10.12300/j.issn.1674-5817.2026.069.

ZHANG Jiaoling,LIN Aping,XU Shiying,et al. Optimization and Application of Mouse Embryo Transfer Technology[J]. Laboratory Animal and Comparative Medicine, 2026, 46(5): 750-758. DOI: 10.12300/j.issn.1674-5817.2026.069.

图/表 4

图1 26G针头打孔与维纳斯显微剪剪切两种输卵管开口方式对ICR假孕小鼠胚胎移植妊娠率、仔鼠出生率的影响

注：A，维纳斯显微剪剪切输卵管开口操作示意图；B，26G注射器针头打孔输卵管开口操作示意图；C，两种开口方式下受体母鼠妊娠率的统计结果；D，两种开口方式下移植仔鼠出生率的统计结果。^*** P<0.001。

Figure 1 Effects of two oviduct opening methods （Vannas scissors cutting <i>vs</i>. 26G needle puncture） on pregnancy rate and pup birth rate of pseudopregnant ICR mice during embryo transfer

Note： A， Schematic diagram of making an opening in the oviduct using Vannas scissors； B， Schematic diagram of making an opening in the oviduct using a 26G syringe needle； C， Statistical results of pregnancy rate in recipient mice with two oviduct opening methods； D， Statistical results of pup birth rate after embryo transfer with two oviduct opening methods. ^*** P<0.001.

图2 不同体重区间ICR假孕受体对胚胎移植妊娠率、仔鼠出生率及7日龄仔鼠存活率的影响

注：A，不同体重区间受体母鼠的妊娠率对比；B，不同体重区间胚胎移植仔鼠的出生率对比；C，不同体重区间出生仔鼠的7 d存活率对比。^* P<0.05，^** P<0.01，^*** P<0.001，^**** P<0.000 1。

Figure 2 Effects of different body weight ranges of pseudo-pregnant ICR recipients on pregnancy rate， pup birth rate， and 7-day pup survival rate following embryo transfer

Note： A， Comparison of pregnancy rate in recipient mice among different body weight groups； B， Comparison of pup birth rate among different body weight groups； C， Comparison of 7-day pup survival rate among different body weight groups. ^* P<0.05， ^** P<0.01， ^*** P<0.001， ^**** P<0.000 1.

图3 单次移植不同数量2细胞胚胎对ICR假孕小鼠妊娠率与仔鼠出生率的影响

注：A，5～15枚、16～25枚、26～35枚、36～45枚大梯度胚胎移植受体的妊娠率；B，5～15枚、16～25枚、26～35枚、36～45枚大梯度胚胎移植受体的仔鼠出生率；C和D，16～35枚细分胚胎数量小梯度胚胎移植受体的妊娠率和仔鼠出生率。^ns P>0.05， ^**P<0.01， ^***P<0.001， ^****P<0.000 1。

Figure 3 Effects of the number of 2-cell embryos transferred per recipient in a single transfer on pregnancy rate and pup birth rate in pseudopregnant ICR mice

Note： A， Pregnancy rates in recipients transplanted with embryos at gradient doses of 5-15， 16-25， 26-35， and 36-45 embryos； B， Pup birth rates in the same four groups； C and D， Pregnancy and pup birth rates， respectively， in recipients grouped by finer embryo-number intervals with the range of 16-35 embryos. ^ns P>0.05， ^** P<0.01， ^*** P<0.001， ^**** P<0.000 1.

图4 单侧输卵管移植与双侧输卵管移植不同手术方式对ICR假孕小鼠妊娠率、仔鼠出生率的影响

注：A，输卵管单侧/双侧移植受体母鼠的妊娠率；B，输卵管单侧/双侧移植仔鼠的出生率。^ns P>0.05。

Figure 4 Effects of unilateral and bilateral oviduct embryo transfer on pregnancy rate and pup birth rate in pseudopregnant ICR mice

Note： A， Pregnancy rate of recipient mice subjected to unilateral or bilateral oviduct embryo transfer； B， Pup birth rate after unilateral or bilateral oviduct embryo transfer. ^ns P>0.05.

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小鼠胚胎移植技术的优化及应用

Optimization and Application of Mouse Embryo Transfer Technology

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