Abstract
Purpose: The objective of this study was to evaluate the effects of growth factor supplementation and Vero cell co-culture on apoptosis and development of frozen thawed one-cell mouse embryos.
Methods: The following treatment regimens were assessed: (a) control medium (b) Vero cell co-culture and (c) growth factor supplemented medium. The individual growth factors tested were: GM-CSF, IGF-I, IGF-II, TNF-α, FGF-4, LIF, TGF-α, TGF-β, IL-6, PDGF and EGF. Blastocyst development and differentiation were monitored. At termination of the experiments, overall blastomere number and apoptosis were assessed using the TUNEL assay.
Results: No differences were observed in blastulation and hatching rates. ICM differentiation in thawed embryos was notably improved with either co-culture or growth factor supplementation. The only growth factor significantly modulating apoptosis in thawed embryos was granulocyte-macrophage colony stimulating factor (GM-CSF). GM-CSF enhanced continued cell survival and prevented apoptosis but did not influence overall cell number in developing blastocysts. Vero cell co-culture significantly increased cell number in blastocysts (124±42 vs 100±44 in control; P<0.05). Embryonic apoptosis was higher in the co-cultured embryos. The increased presence of apoptotic cells in blastocysts of high cell number may reflect the regulatory role of apoptosis in balancing ICM: TE ratios.
Conclusion: These data indicate that culture conditions can modulate post-thaw embryonic development and apoptosis.
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Financial support: Research grant from the Cleveland Clinic Foundation, Cleveland, Ohio, USA.
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Presented in part at: the 60th annual meeting of the American Society for Reproductive Medicine, 2004, October 16–20, Philadelphia, PA, USA.
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Desai, N., Kattal, N., AbdelHafez, F.F. et al. Granulocyte-macrophage colony stimulating factor (GM-CSF) and co-culture can affect post-thaw development and apoptosis in cryopreserved embryos. J Assist Reprod Genet 24, 215–222 (2007). https://doi.org/10.1007/s10815-007-9119-8
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DOI: https://doi.org/10.1007/s10815-007-9119-8