C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size

Embryo transfer (ET) is a common procedure in rodent facilities. Optimizing this technique may help to reduce the number of animals, but little information is available regarding wild type strains and the conditions that affect embryo transfer. To explore this theme, 2-cell C57BL/6J embryos were tra...

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Main Authors: Sofia Lamas, Filipa Franquinho, Marlene Morgado, João R. Mesquita, Fátima Gärtner, Irina Amorim
Format: Article
Language:English
Published: MDPI AG 2020-08-01
Series:Animals
Subjects:
Online Access:https://www.mdpi.com/2076-2615/10/8/1424
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author Sofia Lamas
Filipa Franquinho
Marlene Morgado
João R. Mesquita
Fátima Gärtner
Irina Amorim
author_facet Sofia Lamas
Filipa Franquinho
Marlene Morgado
João R. Mesquita
Fátima Gärtner
Irina Amorim
author_sort Sofia Lamas
collection DOAJ
description Embryo transfer (ET) is a common procedure in rodent facilities. Optimizing this technique may help to reduce the number of animals, but little information is available regarding wild type strains and the conditions that affect embryo transfer. To explore this theme, 2-cell C57BL/6J embryos were transferred after overnight culture of freshly collected zygotes using different conditions: unilateral transfers using a total of 6, 8, 12, 15, 20 and 25 embryos were performed initially; then, this strain was also used for bilateral transfers using a total of 6, 12 and 20 embryos equally divided by the two oviducts. Groups of 25 embryos were not tested for the bilateral technique, since this condition produced the lower success rate when using the unilateral technique and 20 embryos would still represent a large number of embryos. A group of 2-cell B6129F1 embryos was also transferred using unilateral and bilateral ET with 6, 12 and 20 embryos. Crl:CD1(ICR) were used as recipient females for non-reciprocal transfers and C57BL/6J were used to test reciprocal transfers (only tested for six C57BL/6J unilateral transfers). Unilateral transfers using C57BL/6J mice produced higher success rates using six embryos, compared to the other groups transferred unilaterally (<i>p</i>-values between 0.0001 and 0.0267), but the mean number of pups per litter was not different among groups. Bilateral transfer produced higher number of pups when 20 embryos were divided by the two oviducts compared to six (<i>p</i> = 0.0012) or 12 (<i>p</i> = 0.0148) embryos, but with no differences in success rates. No statistical differences were found between the groups of B6129F1, but better results were obtained on bilateral transfers using a total of six embryos. For the strain tested (C57BL/6J), the uterine environment (Crl:CD1(ICR) or C57BL/6J recipient) does not impact the outcome of the technique. These results complement previous work published using genetically engineered mice strains and show that unilateral transfers using low number of embryos (6), produce better outcomes when compared to bilateral or unilateral transfers using more embryos. It also highlights differences between the outcome of bilateral transfers in the two strains tested. A set of historical data of genetically engineered mice at a C57BL/6J background was also included, confirming that lower embryo numbers are related to higher success rates. Together, the outcome of these experiments can be important to reduce the number of recipient and donor females, optimize embryo transfers and improve animal welfare discouraging the use of a more invasive technique.
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spelling doaj.art-c6e8ea08e55742cbbc22bf3beee6a3112023-11-20T10:12:29ZengMDPI AGAnimals2076-26152020-08-01108142410.3390/ani10081424C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter SizeSofia Lamas0Filipa Franquinho1Marlene Morgado2João R. Mesquita3Fátima Gärtner4Irina Amorim5Animal Facility, i3S/ IBMC, Rua Alfredo Allen 208, 4200-135 Porto, PortugalAnimal Facility, i3S/ IBMC, Rua Alfredo Allen 208, 4200-135 Porto, PortugalAnimal Facility, i3S/ IBMC, Rua Alfredo Allen 208, 4200-135 Porto, PortugalEpidemiology Research Unit (EPIUnit), Institute of Public Health, University of Porto, 4050-313 Porto, PortugalAnimal Facility, i3S/ IBMC, Rua Alfredo Allen 208, 4200-135 Porto, PortugalInstitute of Biomedical Science Abel Salazar—ICBAS, R. Jorge de Viterbo Ferreira 228, University of Porto, 4050-313 Porto, PortugalEmbryo transfer (ET) is a common procedure in rodent facilities. Optimizing this technique may help to reduce the number of animals, but little information is available regarding wild type strains and the conditions that affect embryo transfer. To explore this theme, 2-cell C57BL/6J embryos were transferred after overnight culture of freshly collected zygotes using different conditions: unilateral transfers using a total of 6, 8, 12, 15, 20 and 25 embryos were performed initially; then, this strain was also used for bilateral transfers using a total of 6, 12 and 20 embryos equally divided by the two oviducts. Groups of 25 embryos were not tested for the bilateral technique, since this condition produced the lower success rate when using the unilateral technique and 20 embryos would still represent a large number of embryos. A group of 2-cell B6129F1 embryos was also transferred using unilateral and bilateral ET with 6, 12 and 20 embryos. Crl:CD1(ICR) were used as recipient females for non-reciprocal transfers and C57BL/6J were used to test reciprocal transfers (only tested for six C57BL/6J unilateral transfers). Unilateral transfers using C57BL/6J mice produced higher success rates using six embryos, compared to the other groups transferred unilaterally (<i>p</i>-values between 0.0001 and 0.0267), but the mean number of pups per litter was not different among groups. Bilateral transfer produced higher number of pups when 20 embryos were divided by the two oviducts compared to six (<i>p</i> = 0.0012) or 12 (<i>p</i> = 0.0148) embryos, but with no differences in success rates. No statistical differences were found between the groups of B6129F1, but better results were obtained on bilateral transfers using a total of six embryos. For the strain tested (C57BL/6J), the uterine environment (Crl:CD1(ICR) or C57BL/6J recipient) does not impact the outcome of the technique. These results complement previous work published using genetically engineered mice strains and show that unilateral transfers using low number of embryos (6), produce better outcomes when compared to bilateral or unilateral transfers using more embryos. It also highlights differences between the outcome of bilateral transfers in the two strains tested. A set of historical data of genetically engineered mice at a C57BL/6J background was also included, confirming that lower embryo numbers are related to higher success rates. Together, the outcome of these experiments can be important to reduce the number of recipient and donor females, optimize embryo transfers and improve animal welfare discouraging the use of a more invasive technique.https://www.mdpi.com/2076-2615/10/8/1424embryo transferlitter sizemouse unilateral and bilateral surgical transfers
spellingShingle Sofia Lamas
Filipa Franquinho
Marlene Morgado
João R. Mesquita
Fátima Gärtner
Irina Amorim
C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size
Animals
embryo transfer
litter size
mouse unilateral and bilateral surgical transfers
title C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size
title_full C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size
title_fullStr C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size
title_full_unstemmed C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size
title_short C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size
title_sort c57bl 6j and b6129f1 embryo transfer unilateral and bilateral transfer embryo number and recipient female background control for the optimization of embryo survival and litter size
topic embryo transfer
litter size
mouse unilateral and bilateral surgical transfers
url https://www.mdpi.com/2076-2615/10/8/1424
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