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Relationship between age of blastocyst formation and interferon-τ secretion by in vitro–derived bovine embryos
Authors:H Michael Kubisch  Melissa A Larson  R Michael Roberts
Abstract:This study was designed to examine the relationship between the speed at which bovine embryos reach the blastocyst stage, their cell number, and interferon-τ production. A total of 800 oocytes were fertilized by frozen-thawed semen. On day 2, 44 hr after exposure to sperm, 78, 320, and 296 embryos were at the two-, four-, and eight-cell stages, respectively, with an overall cleavage rate of 86.8%. Within these three groups 15 (19.2%), 106 (33.1%), and 158 (53.4%) embryos proceeded to the blastocyst stage. Of these 46.7%, 65.1%, and 63.3% hatched in the three groups, respectively. Blastocysts began to appear at day 7, but a few did not form until as late as day 13. Expanded blastocysts (n = 279) were cultured individually for 48 hr in 50-μl droplets of medium, fixed for cell counts, and the concentration of interferon-τ in the medium was determined. Blastocysts originating from two-cell embryos had significantly fewer cells (46.5 ± 23.3) than either four-cell- (97.2 ± 13.5) or eight-cell-derived embryos (113.8 ± 13.6; P < 0.05). Hatching was accompanied by an increase in cell number (129.8 ± 15.5 versus 41.9 ± 14.4; P < 0.01). Blastocysts derived from embryos that had reached the eight- or four-cell stage 44 hr after insemination produced significantly more interferon than embryos derived from two-cell embryos (941.7 ± 92.1, 930.1 ± 163.1, versus 232.8 ± 70.1 pM). In contrast, hatching, ovary batch, the speed of early cleavage, cell number, and quality grade had no effect on interferon-τ secretion. The embryo's age at blastocyst formation was not related to the number of its cells but did have a significant effect (P < 0.001) on interferon-τ production, with mean concentrations in the medium of 294.8 ± 57.9, 563.3 ± 82.0, 1126.3 ± 133.6, 1778.5 ± 297.2, 512.9 ± 82.0, 315.0 ± 157.5, and 157.5 pM among blastocysts appearing from days 7 to 13, respectively. These data suggest that blastocysts that form at days 7 and 8 produce less interferon-τ than those that form on days 9 or 10. Since early-forming blastocysts are generally considered more developmentally competent than those which form late, there may be a negative relationship between early interferon-τ production and competence. Mol. Reprod. Dev. 49:254–260, 1998. © 1998 Wiley-Liss, Inc.
Keywords:cattle  blastocyst  developmental competence  cleavage rate  in vitro maturation  in vitro fertilization
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