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  • 1
    Electronic Resource
    Electronic Resource
    In vitro preimplantation mouse embryo development with incubation temperatures of 37 and 39°C (1992)
    Springer
    Journal of assisted reproduction and genetics 9 (1992), S. 149-154 
    Details
    ISSN: 1573-7330
    Keywords: embryo culture ; incubation temperatures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Embryos from two strains of mice were used to assess the effect of incubation temperature on pronuclear and twocell development to the morula/blastocyst (M/B) stage. Embryos from B6D2F2 and B6SJLF1 strains were cultured in medium M16 at either 37 or 39°C until 120 hr post human chorionic gonadotropin (hCG) or 0, 24, or 48 hr at 37°C and the remaining time at 39°C. Overall M/B development for pronuclear embryos was 0.6, 0, 32.3, and 52.4% for 0—96, 24—72, 48—48, and 96—0 hr at 37 and 39°C, respectively. Only 0—96 and 24—72 hr at 37 and 39°C were not different (P 〉0.10). Overall M/B development for two-cell embryos was 48.1, 78.1, and 98.0% for 0—72, 24—48, and 72—0 hr at 37 and 39°C, respectively. Percentage development at each time was different (P 〈.01) for each category. Additionally, the number of nuclei for morulae and blastocysts tended to be higher for embryos initiating culture at the two-cell stage compared to pronuclear embryos. The first cell cycle was most dramatically affected by a 2°C increase in incubator temperature. More advanced embryos can tolerate slight increases in incubator temperature more readily than pronuclear embryos.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01203755
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  • 2
    Electronic Resource
    Electronic Resource
    Embryo density and medium volume effects on early murine embryo development (1992)
    Springer
    Journal of assisted reproduction and genetics 9 (1992), S. 454-457 
    Details
    ISSN: 1573-7330
    Keywords: embryo culture ; density ; culture volume
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Methods Embryos were collected from immature C57BL6 female mice superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin and mated by CD1 males. Groups of 1, 5, 10, or 20 embryos were cultured in 5-, 10-, 20-, or 40-µl drops of CZB under silicon oil at 37.5°C in a humidified atmosphere of 5% CO 2 and 95% air.
    Abstract: Results Development score for embryos cultured in 10 µl was higher than that of embryos cultured in 20 or 40 µl. Embryos cultured in groups of 5, 10, or 20 had higher development scores than embryos cultured singly. The highest development score was obtained by the combination of 5 embryos per 10-µl drop. The percentage of live embryos in 20 or 40 µl was lower than that of embryos cultured in 10 µl. Additionally, the percentage of live embryos cultured singly was lower than that of embryos cultured in groups.
    Notes: Background One-cell mouse embryos were used to determine the effects of drop size and number of embryos per drop for optimum development in vitro.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01204051
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  • 3
    Electronic Resource
    Electronic Resource
    Influence of time of gene microinjection on development and DNA detection frequency in bovine embryos (1994)
    Springer
    Transgenic research 3 (1994), S. 226-231 
    Details
    ISSN: 1573-9368
    Keywords: DNA integration ; transgenics ; micromanipulation ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of DNA microinjection at various times afterin vitro insemination on DNA detection and survival rates of bovine embryos was investigated. Oocytes were inseminated 24 h after maturation with frozen/thawed semen prepared with a Percoll separation procedure. At 11, 15 and 19 h after insemination, embryos were centrifuged to visualize pronuclei and microinjected with a murine whey acidic protein-human protein C genomic DNA construct. After culture for 7 days on Buffalo Rat Liver cells, embryos were assessed for stage of development and assayed for the presence of the transgene by polymerase chain reaction. Of zygotes in the 11h after insemination treatment, 16% (25/152) of non-injected and 7% (11/161) of injected embryos developed to the morula or blastocyst stage. Comparable development of non-injected and injected embryos treated at 15h after insemination was 15% (23/158) and 4% (6/159) and treated at 19 h after insemination was 14% (23/162) and 1% (1/165), respectively. Development of injected embryos was greater (p〈0.05) when injection was performed at 11 h after insemination compared to 19 h after insemination. Development of non-injected embryos was greater (p〈0.01) than that of injected embryos. There was no difference in transgene detection frequency in embryos of all developmental states between treatments (53% at 11; 50% at 15; 48% at 19h after insemination). Injected embryos testing positive for the presence of the transgene exhibited increased development over negative embryos (p〈0.01). Greater development efficiencies can be obtained in microinjected bovine embryos when injection is performed early in pronuclear formation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02336775
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  • 4
    Electronic Resource
    Electronic Resource
    Gene transfer efficiency during gestation and the influence of co-transfer of non-manipulated embryos on production of transgenic mice (1994)
    Springer
    Transgenic research 3 (1994), S. 20-25 
    Details
    ISSN: 1573-9368
    Keywords: Gene transfer ; gestational losses ; non-manipulated mice embryos
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Litter size of DNA microinjected zygotes is lower than for non-manipulated zygotes. The rate of embryonic and fetal survival in early, mid and late gestation was determined to assess whether DNA integration was responsible for embryonic losses. Also, the effect of including non-microinjected embryos with injected embryos on pregnancy rate and transgenic pup production was determined. In Experiment 1, one-cell embryos from immature CD-1 mice were microinjected with a whey acidic protein promoter-human protein C gene construct. One hour after microinjection embryos were transferred to pseudopregnant recipients (45 transfers of 30 embryos each). Fifteen recipients were sacrificed on day 4, 12 and 18 of gestation and the embryos/fetuses analysed for the transgene. The percentage of embryos or fetuses that were positive for the transgene was not significantly different at any day. However, the number of viable embryos at day 4 was significantly greater than fetuses on days 12 or 18. In addition, a high degree of mosaicism was observed in day 18 fetuses and placentae recovered. In Experiment 2, one-cell embryos from CD-1 mice were microinjected and co-transferred with non-manipulated embryos (C57BL/6). Pregnancy rate and the total number of pups born were improved by addition of non-injected embryos. However, the number of transgenic mice produced was similar whether non-injected embryos were included or not. There were 32.2% (15/46) transgenic pups when 0 non-injected embryos were transferred compared with 15.1% (13/86) transgenic pups when 4 or 8 non-injected embryos were added to the transfers. In summary, a high degree of embryonic and fetal mortality occurs among microinjected embryos. Furthermore, since the percentage of transgenesis did not change throughout pregnancy, DNA integration does not appear to account for all of the embryonic losses. other factor(s) related to the microinjection procedure may be involved in the embryonic and fetal failure of microinjected embryos. Addition of non-injected embryos, although it increased pregnancy rate and the number of pups born from microinjected embryos, actually decreased the number of transgenic pups obtained per pregnancy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01976023
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    Paper (German National Licenses)
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