J. Anim Sci.
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Journal of Animal Science, Vol 75, Issue 5 1332-1336, Copyright © 1997 by American Society of Animal Science

JOURNAL ARTICLE

Culture of bovine embryos in buffalo rat liver cell-conditioned media or with leukemia inhibitory factor

R. N. Funston, W. J. Nauta and G. E. Seidel Jr
Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins 80523, USA.

Experiments were designed to study development of bovine embryos in TCM-199 medium conditioned by preculture with buffalo rat liver (BRL) cells. Conditioned media were harvested after BRL cells were cultured until confluency (CON), or for an additional 2 d with the same cells but new medium (CON-N) or the same medium (CON-S). Glucose in TCM-199 was depleted by BRL cells to different concentrations depending on coculture procedures: CON = 3.94 mM, CON-N = 1.67 mM, and CON-S = 1.11 mM glucose. In Exp. 1, development of bovine zygotes in CON-S resulted in fewer blastocysts than development in CON (10 vs 28%, P < .05); CON-N was not different from CON (26% blastocysts). Experiment 2 examined effects of moving embryos to a fresh drop of different or identical conditioned medium after culture for 3 d. Initial culture in CON-N and final culture in CON resulted in a greater (P < .01) number of blastocysts compared with the control of CON followed by CON (32 vs 19% blastocysts). This was not entirely due to changing from low to high glucose because adding glucose to CON-N after 3 d yielded only 18% blastocysts. To test the hypothesis that beneficial effects of BRL cell-conditioned media may be due to secretion of leukemia inhibiting factor (LIF), LIF was added to B2, a more appropriate medium than Medium-199 for culturing bovine embryos without conditioning or coculture with BRL cells. In the absence of serum, the percentage of blastocysts per cleaved embryo (17 to 28%) was not improved with LIF; however, the mean number of cells per blastocyst was higher (P < .05) in treatments with LIF (65 to 74 cells) than without LIF (47 cells). In B2 medium + 10% fetal calf serum, LIF was of no benefit; development to blastocysts was good with or without LIF (43% of cleaved).


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G-P Li, K L White, K I Aston, L N Meerdo, and T D Bunch
Conditioned medium increases the polyploid cell composition of bovine somatic cell nuclear-transferred blastocysts
Reproduction, February 1, 2004; 127(2): 221 - 228.
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Copyright © 1997 by the American Society of Animal Science.