Optimization of embryo transfer protocols for mice

Johnson, Larry; Moffatt, R. J.; Bartol, Frank F.; Pinkert, Carl A.

doi:10.1016/s0093-691x(96)00298-1

Cited by 21 publications

(27 citation statements)

References 9 publications

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“…Zygotes can be routinely cultured to the two-cell or blastocyst stage, then transferred to the oviduct of day 1 pseudopregnant mice (Bronson and McLaren, 1970;Kooyman et al, 1990;Johnson et al, 1996). Using outbred (e.g., ICR) or hybrid (e.g., B6SJLF1) mice, typically 40À60% and 60À80% of injected eggs (60À80 and 75À100% of noninjected eggs), respectively, develop overnight to the two-cell stage in our experience.…”

Section: In Vitro Culture Of Microinjected Eggsmentioning

confidence: 81%

“…Several different hybrids are currently popular for DNA microinjection; the C57BL/6 3 SJL (B6SJL) F1 hybrid has been shown to be efficient in the generation of transgenic mice (Brinster et al, 1985;Johnson et al, 1996;Vergara et al, 1997). The majority of the hybrids used for microinjection utilize the C57BL/6 inbred strain as one of the parental stocks due to favorable genetic and embryological characteristics.…”

Section: A Strainsmentioning

confidence: 99%

“…In experiments where other strains are utilized, losses in experimental efficiencies will be encountered. In some instances (e.g., using FVB or C57BL/6 strains), efficiency losses may be minimal, but conditions for manipulation and timing of specific biological end points (e.g., pronuclear egg formation, response to gonadotropins) using various strains must be identified (Johnson et al, 1996;Vergara et al, 1997). As with strains used in different fields (e.g., other inbreds or congenics), efficiency losses can range from a few percent to more than a 100-fold difference (using DBA/2 congenic mice as compared to the B6SJLF1 hybrid).…”

Section: A Strainsmentioning

confidence: 99%

“…One or two control eggs may be transferred to help ensure pregnancy maintenance if there is concern regarding proficiency or quantity of microinjected eggs, as just two implanted eggs have been shown to be as effective at maintaining pregnancy as larger numbers Johnson et al, 1996. ) Alternatively, eggs may be deposited directly into the ampulla via a previously made hole or cut between the ostium and ampulla, thus avoiding trouble arising from anatomical variations often found in the ostium (Johnson et al, 1996). The reproductive tract is then carefully replaced into the abdomen, and the body wall is closed with one or two sutures followed by skin closure with wound clips.…”

Section: Ova Reimplantationsmentioning

confidence: 99%

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DNA Microinjection, Embryo Handling, and Germplasm Preservation

Polites

Johnson

Pinkert

2014

Transgenic Animal Technology

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Section: In Vitro Culture Of Microinjected Eggsmentioning

confidence: 81%

Section: A Strainsmentioning

confidence: 99%

Section: A Strainsmentioning

confidence: 99%

Section: Ova Reimplantationsmentioning

confidence: 99%

See 2 more Smart Citations

DNA Microinjection, Embryo Handling, and Germplasm Preservation

Polites

Johnson

Pinkert

2014

Transgenic Animal Technology

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“…In addition to the well-known protocols for oviduct and uterus transfer published by Nagy et al [9], some variations have been described, such as uterine transfer via the uterotubal junction [10] or puncture of the oviductal wall [11,12]. In all these protocols, recipients were used only once.…”

Section: Introductionmentioning

confidence: 99%

Repeated Use of Surrogate Mothers for Embryo Transfer in the Mouse

Kolbe

Palme

Touma

et al. 2012

Biology of Reproduction

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Embryo transfer in mice is a crucial technique for generation of transgenic animals, rederivation of contaminated lines, and revitalization of cryopreserved strains, and it is a key component of assisted reproduction techniques. It is common practice to use females only once as surrogate mothers. However, their reuse for a second embryo transfer could provide hygienic and economic advantages and conform to the concept of the 3Rs (replace, reduce, refine). This investigation evaluated the potential for a second embryo transfer in terms of feasibility, reproductive results, and experimental burden for the animal. Virgin female ICR mice (age 8-16 wk) were used as recipients for the first embryo transfer. Immediately after weaning of the first litter, a second surgical embryo transfer was performed into the same oviduct. Virgin females of comparable age to the reused mothers served as controls and underwent the same procedure. The first surgery did not affect the success of the second embryo transfer. Histological sections showed excellent wound healing without relevant impairment of involved tissues. We observed no differences in pregnancy rates or litter sizes between the transfer groups. Most importantly, we found no change in behavior indicating reduced well-being and no increase of corticosterone metabolites in the feces of surrogate mothers reused for a second embryo transfer. We conclude that a second embryo transfer in mice is feasible with regard to reproductive and animal welfare aspects.

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Effect of chemically defined culture medium supplemented with β‐mercaptoethanol and amino acids on implantation and development of different stage in vivo‐ or in vitro‐derived mouse embryos

Bağış

Mercan

2004

Molecular Reproduction Devel

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In vitro culture (IVC) systems are required for many biotechnological and assisted reproductive technologies and the researchers have been modifying in vitro embryo culture conditions to reach the comparable efficiencies provided in vivo. In the present study, the effects of beta-mercaptoethanol (Beta-ME) and amino acids (AA) on the development of mouse embryos obtained in vivo or in vitro at different stages were investigated. Chemically defined potassium simplex optimized medium (KSOM) was used as basic culture medium and six experimental groups were established and by supplementation of Beta-ME and AA into KSOM media. The quality of blastocysts was evaluated by counting the cells and determining the ratio of inner cell mass (ICM) to trophoectoderm (TE) cells. In addition, embryo transfer (ET) was performed to investigate the rate of implantation and live fetuses. The results obtained in the present study demonstrated that the combined treatment of Beta-ME and AA to 1-cell stage embryos not only enhanced in vitro development to the blastocyst stage but also improved both the number of blastocysts cells and live fetuses.

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Optimization of embryo transfer protocols for mice

Cited by 21 publications

References 9 publications

DNA Microinjection, Embryo Handling, and Germplasm Preservation

DNA Microinjection, Embryo Handling, and Germplasm Preservation

Repeated Use of Surrogate Mothers for Embryo Transfer in the Mouse

Effect of chemically defined culture medium supplemented with β‐mercaptoethanol and amino acids on implantation and development of different stage in vivo‐ or in vitro‐derived mouse embryos

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