This study focuses on the effects of pentoxifylline and 2-deoxyadenosine, commonly used in human assisted reproduction, on mouse embryos development.
Female mice of ICR strain (5-6 weeks old) were injected with 5IU of PMSG and hCG 46-48 hours intervals, and then were caged with fertile male. 2-cell embryos were obtained by flushing the oviducts at 45-46 hours post hCG injection. Collected
embryos
were cultured in vitro for 96 hours by using microdroplet method at 37℃ in an incubator supplied with 5% CO2 in air and 100% humidity.
In experiment 1, 2-cel embryos were cultured in vitro in medium containing 3mM pentoxifylline and 3mM 2-deoxyadenosine for 96 hours. In experiment 2, 2-cell embryos were incubated for 30 minutes in medium containing 3mM pentoxifylline and 3mM
2-deoxy-adenosine. Then, 2-cell embryos were washed and cultured in control medium (M16+0.4% BSA). Embryos were observed daily and classified as 4-8 cell, morulae, blastocyst stage. Cell numbers of blastocysts were counted according to Takowski's
method.
In experiment 1, 3mM pentoxifylline inhibited the development of morulae and blastocysts. At 3mM 2-deoxyadenosine inhibited cleavage from the 4-8 cell stage onwards.
In experiment 2, when 2-cell mouse embryos were exposed for 30 minutes to 3mM pentoxifylline and 3mM 2-deoxyadenosine, blastocyst formation was morphologically normal. However, cell numbers after exposure to pentoxifylline and 2-deoxyadenosine
were
significantly decreased in comparison to embryos in the control group. There was, however, no difference between the cell numbers of embryos exposed to 3mM pentoxifylline or 3mM 2-deoxy-adenosine.
From these results, we know that 3mM pentoxifylline and 3mM 2-deoxyadenosine might be detrimental for embryonic development of mouse. Therefore, exposure of the early embryos to pentoxifylline and 2-deoxyadenosine should be avoided as possible,
when
this drugs are used in human assisted reproduction. As mechanisms of embryo-toxicity of pentoxifylline and 2-deoxyadenosine are unclear, futher research is needed in this field.
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