Abstract
Growth factors and cytokines are present in small quantities in the oviduct and uterus and some are synthesized by the growing embryo. Granulocytes-macrophage colony-stimulating factor (GM-CSF) is known as an important regulator, which enhances cell proliferation and reduces apoptosis in developing blastocysts, during normal fetal and placental development. The purpose of this study is to investigate whether adding GM-CSF to the culture media affects blastulation or the chromosomal status of mouse embryos.
Murine embryos were cultured in vitro from the 2-cell stage until the blastocyst stage in the presence of different concentrations of GM-CSF of 0 ng/ml (control), 1, 2, 5 and 10 ng/ml. The development of each embryo was noted and the embryos were then spread for fluorescence in situ hybridization (FISH) using locus-specific probes (LSI) for chromosomes 2, 11 and 16 in all embryos.
No difference in the blastulation potential was noted with the addition of 1 and 2 ng/ml of GM-CSF compared with the controls, but there was a significant decrease (P < 0.001) in the blastulation rate in the 5 and 10 ng/ml concentrations. The rate of mosaicism/aneuploidy noted in all GM-CSF groups (1, 2, 5 and 10 ng/ml) was slightly higher than in the control group (0 ng/ml GM-CSF) but the differences were not significant. In the mosaic embryos from the GM-CSF cultured groups, the percentage of aneuploid cells was statistically higher than in the control group.
GM-CSF exerted a negative impact on blastocyst development at higher concentrations. GM-CSF did not affect the rates of mosaicism/aneuploidy, but did increase the percentage of aneuploid cells within the mosaic embryos. Adding GM-CSF to the culture media for clinical use requires further studies either on human or animal models to evaluate its long-term effects.