“…The role of hLIF in bovine embryo culture has been controversial in previous studies [8][9][10][11] in which the effects of different supplements, such as serum, synthetic molecules and BSA were examined. In the present study, embryonic development was impaired by hLIF, while the number of ICM cells was reduced by mLIF, in disagreement with the effects described by Yamanaka et al for bLIF [6,7].…”
Section: Discussionmentioning
confidence: 99%
“…When added from the morula stage, hLIF has been reported to have no effect on subsequent development [8,9]. During the course of blastocyst formation, hLIF has been found to stimulate expansion and hatching [8,10], but other authors report no comparable effect [11]. It has also been described that the number of cells in the inner cell mass (ICM) and total cell numbers increase in the presence of hLIF, although this led to no improvement in pregnancy and survival rates to cryopreservation [8,9].…”
Leukemia inhibitory factor (LIF) is a cytokine that shows conflicting effects on in vitro produced (IVP) bovine embryos. Bovine LIF (bLIF) has been cloned and used in culture, but there is no commercially available bLIF. Thus, researchers use human LIF (hLIF) to supplement the culture medium for bovine embryos because of its greater sequence homology compared to murine LIF (mLIF). We compared the effects of mLIF and hLIF on the development of bovine embryos in culture with the effects described for bLIF. Oocytes were matured and fertilized in vitro and cultured in modified synthetic oviduct fluid with BSA. On Day 6 postinsemination, morulae were cultured for 48 h in the presence of: (1) mLIF, 100 ng ml À1 ; (2) hLIF, 100 ng ml À1 ; or (3) no LIF. Reduced blastocyst rates were observed on Day 8 for hLIF at the middle and expanded stages, while mLIF had no effect. In contrast, Day 8 blastocysts showed decreased cell counts both in terms of inner cell mass (ICM) and ICM/total cell proportions in the presence of mLIF, while hLIF had no effect. No changes were seen in trophectoderm (TE) and total cell counts. The increased hatching rates and TE cell counts previously described for bLIF, together with the disparate effects exhibited by hLIF and mLIF during blastocyst formation indicate these compounds are inappropriate to replace bLIF. We recommend that heterospecific LIF should not be used to supplement the culture medium for bovine embryo or embryonic stem cells.
“…The role of hLIF in bovine embryo culture has been controversial in previous studies [8][9][10][11] in which the effects of different supplements, such as serum, synthetic molecules and BSA were examined. In the present study, embryonic development was impaired by hLIF, while the number of ICM cells was reduced by mLIF, in disagreement with the effects described by Yamanaka et al for bLIF [6,7].…”
Section: Discussionmentioning
confidence: 99%
“…When added from the morula stage, hLIF has been reported to have no effect on subsequent development [8,9]. During the course of blastocyst formation, hLIF has been found to stimulate expansion and hatching [8,10], but other authors report no comparable effect [11]. It has also been described that the number of cells in the inner cell mass (ICM) and total cell numbers increase in the presence of hLIF, although this led to no improvement in pregnancy and survival rates to cryopreservation [8,9].…”
Leukemia inhibitory factor (LIF) is a cytokine that shows conflicting effects on in vitro produced (IVP) bovine embryos. Bovine LIF (bLIF) has been cloned and used in culture, but there is no commercially available bLIF. Thus, researchers use human LIF (hLIF) to supplement the culture medium for bovine embryos because of its greater sequence homology compared to murine LIF (mLIF). We compared the effects of mLIF and hLIF on the development of bovine embryos in culture with the effects described for bLIF. Oocytes were matured and fertilized in vitro and cultured in modified synthetic oviduct fluid with BSA. On Day 6 postinsemination, morulae were cultured for 48 h in the presence of: (1) mLIF, 100 ng ml À1 ; (2) hLIF, 100 ng ml À1 ; or (3) no LIF. Reduced blastocyst rates were observed on Day 8 for hLIF at the middle and expanded stages, while mLIF had no effect. In contrast, Day 8 blastocysts showed decreased cell counts both in terms of inner cell mass (ICM) and ICM/total cell proportions in the presence of mLIF, while hLIF had no effect. No changes were seen in trophectoderm (TE) and total cell counts. The increased hatching rates and TE cell counts previously described for bLIF, together with the disparate effects exhibited by hLIF and mLIF during blastocyst formation indicate these compounds are inappropriate to replace bLIF. We recommend that heterospecific LIF should not be used to supplement the culture medium for bovine embryo or embryonic stem cells.
“…The addition of human LIF (hLIF) to the culture medium increased the number of eight-cell stage mouse embryos which hatched and increased the size of blastocyst outgrowths in vitro (Robertson et al, 1991;Lavranos et al, 1995). Similarly, the culture of embryos in the presence of hLIF resulted in fewer degenerate embryos and a greater proportion of hatched blastocysts in the sheep (Fry et al, 1992) and cow (Fukui and Matsuyama, 1994;Han et al, 1995). Furthermore (Li and Foote, 1993).…”
Section: Introductionmentioning
confidence: 99%
“…The twocell with a few four-cell embryos were cultured for 3 days (d4 of development) with the exception of those in experiment 3, which were cultured for 5 days (d6 of development). Sixteencell (Fukui and Matsuyama, 1994;Han et al, 1995;Lavranos et al, 1995). Furthermore, the culture interval was increased such that all embryos were cultured until d6 of development, in an effort to discern any effects of hLIP on blastocyst hatching, as had been reported for embryos of the mouse (Robertson et al, 1991;Lavranos et at, 1995), sheep (Fry et al, 1992) and cow (Han et al, 1995).…”
“…During the he technique of in-vitro embryo production is useful tool for the mass production of embry-T os to produce beef calves and to offer materials for basic research in developmental biology. It has been tried to improve the in-vitro developmental potential during preimplantation period by coculture with somatic cells, such as cumulus cells [1], bovine oviductal epithelial cells [2], or buffalo rat liver cells [3], by addition of transforming growth factor [4], leukemia inhibitory factor [5], and epi-MZT, loss or decay of maternal mRNA molecules, activation of transcription from the embryonic genome and marked qualitative changes in protein synthesis in a stage-specific manner occur in mammalian embryos [8].…”
Abstract. RNA synthesis in bovine embryos produced in vitro was investigated to clarify changes in transcriptional activity. Overall 1003 follicular oocytes were matured and fertilized in vitro and subsequently cultured with a cumulus cell monolayer in TCM 199 with 5% fetal bovine serum (FBS). Embryos were transferred to TCM199 supplemented with 100 µCi/ml of [5-3 H]uridine at 26, 35, 43, 50, 68 and 116 h post insemination (hpi) and cultured for 8 h at 39 C. Treated embryos were washed in PBS and the zona pellucida weakened with 5% proteinase K followed by fixation in acetic-alcohol. Preparations were air-dried, dipped in K5 emulsion, and exposed at 4 C. After 4.5 days exposure, the preparations were developed in D19 and the density of 3 H-uridine incorporation was analyzed by image analysis (NIH-157 analysis system). 3 H-Uridine incorporation into nuclei was observed in 43% of 2-cell embryos at 26 hpi although the level was low. The change of 3 Huridine incorporation level was not consistent until 4-cell stage and thereafter the level increased in a time-dependent manner but not dependent on cell number. Incorporation of 3 H-uridine into nucleoli was observed from 50 hpi. Twenty-two embryos at 5-cell to morula were incubated with labelled and unlabelled uridine as negative control showed no incorporation of 3 H-uridine. These results show that the transcription of early bovine embryos fertilized in vitro is already active in 2-cell embryos at 26 hpi, the level increases in a time-dependent manner after 5-cell stage and is not dependent on the cell number.
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