Date of Award
Doctor of Philosophy (PhD)
School of Animal Science
Robert A. Godke
In Experiment I, 125 two-pronuclear murine ova were cultured in either Ham's F-10 plus l5% human serum (HF-10h), on a monolayer of human uterine fibroblasts (HUC) or fetal bovine uterine fibroblasts (FBUC). After 96 hours in culture, 2.4% embryos in HF-10h were viable compared with 45.2% and 61% for embryos on HUC and FBUC, respectively. Results suggest that monolayers were advantageous to medium alone for culturing murine embryos. Experiment II was conducted to assess the development of 117 bovine embryos in HF-10 (Treatment A), on a monolayer of FBUC (Treatment B) or hormone-pretreated FBUC (Treatment C). Embryo viability assessed at 12, 24, 36, 48, 60 and 72 hours in culture. The viability at 72 hours was 38.5%, 87.2% and 76.9% for Treatments A, B and C, respectively. These results indicate FBUC were superior to that of HF-10 medium alone for culturing bovine embryos. In Experiment III, 45 equine embryos were cultured in either Ham's F-12 plus 10% FCS (HF-12), on a monolayer of fetal equine uterine fibroblasts (FEUC) or FBUC. Results suggest that early-stage embryos benefited from the FBUC while later-stage embryos benefited when cultured in HF-12 alone. In Experiment IV, 35 equine embryos were either transferred to recipients or cultured on FBUC for 24 hours prior to transfer to recipients. Pregnancy rate (PR) for fresh embryos was 53.3% and not different from the 42.1% PR for cultured embryos. Results suggest that FBUC could be used as a method for transporting equine embryos. In Experiment V, 272 human IVF zygotes were cultured for at least 24 hours in either Earle's Balanced Salt solution (EBSS) or on FBUC prior to embryo replacement. Incidence of implantation for embryos cultured in EBSS alone was 17% which was lower than the 35% for co-cultured embryos. Results suggest that culture of human zygotes on FBUC could be considered for improving pregnancy rates from human IVF procedures. Overall, fetal cells were advantageous over control media in each experiment in this study. Therefore, one should not overlook this system in future embryo research and commercial opportunities.
Wiemer, Klaus Enrique, "The Development of a Fetal Uterine Cell in Vitro Coculture System for Mammalian Embryos." (1989). LSU Historical Dissertations and Theses. 4823.