Success of murine embryo transfer increased by a modified transfer pipette.
ABSTRACT Embryos transfer is one of the most critical steps for generating genetically modified mice. Because of the limitations of the current transfer equipment and techniques, such as discharge of freshly transferred embryos and the necessity of sophisticated transfer skills, the success ratio of implantation may not well satisfy the requirements for mass production of high quality animals in the field of genetically modified mice. In this study, we describe a modified embryo transfer pipette with a syringe-like tip that can easily be applied to mouse embryo transfer. Using this improved method, we show that the procedures for mouse embryo implantation are simplified and easier to perform; moreover, the viability rate of mouse embryos is 20% higher than that achieved with conventional methods. Our modified tool and improved transfer technique are effective, time-saving and less invasive, resulting in increased success of embryo transfer.
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ABSTRACT: The objective of this study was to evaluate the effect on litter size of 2 analgesics used perioperatively during mouse embryo transfer surgery. Day 2.5 pseudopregnant CD1 mice (n = 96) were divided equally into 2 analgesic treatment groups and a saline control group. Each mouse received a single, subcutaneous dose of buprenorphine hydrochloride (0.1 mg/kg), flunixin meglumine (2.5 mg/kg), or saline immediately after induction of anesthesia with 2.5% isoflurane. Each mouse then was prepared for aseptic surgery. Blastocysts had previously been collected from C57BL/6NCrl female mice that were synchronized and superovulated by using pregnant mare serum gonadotropin and human chorionic gonadotropin and mated with C57BL/6NTac male mice 3.5 d before collection. Viable blastocysts were pooled, and 8 were selected arbitrarily and transplanted into the right uterine horn of each pseudopregnant CD1 mouse. Mice were monitored throughout pregnancy, and the number of pups at birth was documented. No statistically significant difference was found between the 3 groups. These results indicate that perioperative analgesic treatment with buprenorphine or flunixin in the CD1 mouse undergoing embryo transfer is not associated with increased embryonic loss.Journal of the American Association for Laboratory Animal Science: JAALAS 07/2010; 49(4):423-6. · 1.15 Impact Factor
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ABSTRACT: Successful Embryo Transfer (ET) technique is a fateful step of all efforts to achieve live births from in vitro produced embryos in assisted reproductive techniques or in knockout, transgenic or cloned animal projects. Small reproductive tract of mice and limitation of current techniques may not well satisfy the requirements for mass production of genetically modified mice. Genetic abnormalities of embryos, receptivity and uterine contractions, expulsion of embryos, blood, mucus or bacterial contamination on the transfer pipette tip, technical problems and even animal strain may affect embryo transfer outcome. In this study, two techniques of embryo transfer in mice were compared. In conventional technique the oviduct wall was punctured with a 30-gauge needle and the loaded Pasteur pipette with embryos and medium was inserted into the hole. In new technique, embryos that were loaded in modified micropipette with minimal medium were transferred directly to the oviduct by manual piston micro-pump easily. Embryo viability was evaluated considering the percentage of live healthy newborns. Results of the two techniques were compared by t-test within the NPAR1WAY procedure of SAS software (ver. 9.2). The average live birth rates in the novel methods was significantly higher (42.4%) than the conventional method (21.7%, p<0.05). In conclusion, using new embryo transfer technique improved birth rate by preventing embryos expulsion from the oviduct, saving time and easy transfer of embryos with minimum volume of medium.Avicenna journal of medical biotechnology. 01/2013; 5(1):62-5.