Comparison of the interferon-tau expression from primary trophectoderm outgrowths derived from IVP, NT, and parthenogenote bovine blastocysts.
The expression of interferon-tau (IFN-tau) is essential for bovine embryo survival in the uterus. An evaluation of IFN-tau production from somatic cell nuclear transfer (NT)-embryo-derived primary trophectoderm cultures in comparison to trophectoderm cultured from parthenogenote (P) and in vitro matured, fertilized, and cultured (IVP) bovine embryos was performed. In Experiment 1, the success/failure ratio for primary trophectoderm colony formation was similar for IVP and NT blastocysts [IVP = 155/29 (84%); NT 104/25 (81%)], but was decreased (P = .05) for P blastocysts [54/43 (56%)]. Most trophectoderm colonies reached diameters of at least 1 cm within 3-4 weeks, and at this time, 72 hr conditioned cell culture medium was measured for IFN-tau concentration by antiviral activity assay. The amount of IFN-tau produced by IVP-outgrowths [4311 IU/mL (n = 155)] was greater (P < .05) than that from NT- [626 IU/mL (n = 104)] and P - [1595 IU/mL (n = 54)] derived trophectoderm. Differential expression of IFN-tau was confirmed by immunoblotting. In Experiment 2, colony formation was again similar for IVP and NT blastocysts [IVP = 70/5 (93%); NT 67/1 (99%)] and less (P < .05) for P blastocysts [65/27 (70%)]. Analysis of trophectoderm colony size after 23 days in culture showed a similar relationship with P-derived colonies being significantly smaller in comparison to IVP and NT colonies. A differential expression of IFN-tau was also observed again, but this time as measured over time in culture. Maximal IFN-tau production was found at day-14 of primary culture and diminished to a minimum by the 23rd day.
Talbot
Neil C
NC
USDA, ARS, ANRI, Biotechnology and Germplasm Laboratory, Beltsville, Maryland 20705-2350, USA. ntalbot@anri.barc.usda.gov
Powell
Anne M
AM
Ocón
Olga M
OM
Caperna
Thomas J
TJ
Camp
Mary
M
Garrett
Wesley M
WM
Ealy
Alan D
AD
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002417:Cattle; D002467:Cell Nucleus / Q000502:physiology; D003114:Colony-Forming Units Assay; D004475:Ectoderm / Q000502:physiology*; D005260:Female; D005306:Fertilization; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D007370:Interferon Type I / Q000235:genetics*; D009924:Organ Culture Techniques; D010312:Parthenogenesis; D011257:Pregnancy Proteins / Q000235:genetics*; D014327:Trophoblasts / Q000166:cytology / Q000502:physiology*
D007370:Interferon Type I; D011257:Pregnancy Proteins; C020947:interferon tau