小鼠胚胎移植技术的优化及应用

doi:10.12300/j.issn.1674-5817.2026.069

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SPF级ICR小鼠和C57BL/6J小鼠为实验对象，通过体外受精获得优质二细胞胚胎，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28～35 g的假孕受体移植效果最优，妊娠率90%、出生率44%，且无幼崽被母鼠压亡的现象；单只受体移植胚胎数量在16～25枚组出生率达42%为各组最高，26～35枚组妊娠率达90%，且胚胎利用率达到最大化，而胚胎数超过35枚后移植效率显著下降；单只受体胚胎移植数量在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28～35 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: 小鼠, 胚胎移植, 体外受精, 技术优化, 移植效率

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 was used as the evaluation index), and enhance the utilization rate of high-quality embryo germplasm resources(the birth rate of mice after embryo transfer was used as the evaluation index). Methods ICR mice and C57BL/6J mice were used as experimental materials. High-quality 2-cell embryos were obtained through in vitro fertilization. We adopted the single-factor controlled variable method 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 oviduct ostia punctured using a 26G syringe needle reached 90%, which was significantly higher than the 43% in the group with oviduct openings cut by Venus scissors (p<0.05). Pseudopregnant recipients weighing 28-35 g exhibited optimal embryo transfer outcomes, with a pregnancy rate of 90% and a birth rate of 44%, and no pups were crushed by foster mothers. The highest birth rate 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 could balance 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 28-35 g, adopting the oviduct opening method with a 26G syringe needle for puncture, 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: Mouse, Embryo transfer, In vitro fertilization, Technical optimization, Transfer efficiency

中图分类号:

Q95-33

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.. 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. DOI: 10.12300/j.issn.1674-5817.2026.069.

图/表 4

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统计结果；D，两种开口方式下移植仔鼠出生率统计结果。^***p<0.001。
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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、仔鼠出生率的对比
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统计结果；D，两种开口方式下移植仔鼠出生率统计结果。^***p<0.001。

Figure 1 Comparison of two oviduct incision methods （Vannas scissors cutting vs 26G needle puncture） on pregnancy rate and pup birth rate of pseudopregnant ICR mice during embryo transfer
Note： A， Schematic diagram of oviduct ostium incision using Vannas micro-scissors. B， Schematic diagram of oviduct ostium puncture using a 26G syringe needle. C， Statistical results of pregnancy rate in recipient mice with two incision methods. D， Statistical results of pup birth rate after embryo transfer with two incision methods. ^***p < 0.001.

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

Figure 2 Effects of ICR pseudopregnant recipients with different body weight ranges on pregnancy rate， pup birth rate and 7-day-old 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.

注：A和C，5～15枚、16～25枚、26～35枚、36～45枚大梯度胚胎移植受体妊娠率；B和D，对应各组胚胎移植仔鼠出生率；C和D，20～35枚细分胚胎数量梯度结果。^nsp>0.05， ^**p<0.01， ^***p<0.001， ^****p<0.000 1。
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图3 单次移植不同数量二细胞胚胎对ICR假孕小鼠妊娠率与仔鼠出生率的影响
注：A和C，5～15枚、16～25枚、26～35枚、36～45枚大梯度胚胎移植受体妊娠率；B和D，对应各组胚胎移植仔鼠出生率；C和D，20～35枚细分胚胎数量梯度结果。^nsp>0.05， ^**p<0.01， ^***p<0.001， ^****p<0.000 1。

Figure 3 Effects of transferring different numbers of 2-cell embryos in a single operation on pregnancy rate and pup birth rate of pseudopregnant ICR mice
Note： A and C， Pregnancy rates of recipients transplanted with embryos at gradient doses of 5-15， 16-25， 26-35 and 36-45 embryos； B and D， Corresponding pup birth rates in each group. C and D， Outcomes of refined gradient groups with 20-35 embryos. ^nsp>0.05， ^**p<0.01， ^***p<0.001， ^****p<0.000 1.

注：A，输卵管单/双侧移植受体母鼠妊娠率；B，输卵管单/双侧移植仔鼠出生率。^nsp>0.05。
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图4 单侧输卵管移植与双侧输卵管移植不同移植手术方式对ICR假孕小鼠妊娠率、仔鼠出生率影响
注：A，输卵管单/双侧移植受体母鼠妊娠率；B，输卵管单/双侧移植仔鼠出生率。^nsp>0.05。

Figure 4 Comparison of pregnancy rate and pup birth rate between unilateral and bilateral oviduct embryo transfer in pseudopregnant ICR mice
Note： A， Pregnancy rate of recipient mice subjected to unilateral or bilateral oviduct transfer； B， Pup birth rate after unilateral or bilateral oviduct transfer.^nsp>0.05.

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