Prematuration with cyclic adenosine monophosphate modulators alters cumulus cell and oocyte metabolism and enhances developmental competence of in vitro-matured mouse oocytes.

Oocyte in vitro maturation (IVM) is an important assisted reproductive technology and research tool. The adoption of IVM into routine clinical practice has been hindered by its significantly lower success rates compared to conventional in vitro fertilization. Cyclic AMP (cAMP) modulation and follicle-stimulating hormone (FSH), independently, have long been known to improve IVM oocyte developmental competence. This study comprehensively examined the effects of FSH and cAMP/cGMP modulation, alone and in combination, on IVM oocyte metabolism and developmental outcomes. Mouse cumulus-oocyte complexes (COCs) were subjected to a 1 h prematuration phase ± the cAMP modulator forskolin and cAMP/cGMP modulator 3-isobutyl-1-methylxanthine followed by IVM ± FSH. Prematuration with these cyclic nucleotide modulators or IVM with FSH significantly improved oocyte developmental competence and reduced spindle abnormalities compared to spontaneous IVM (no treatment); however, these two treatments in combination endowed even greater developmental competence (improved subsequent blastocyst rates and quality; P < 0.05), albeit blastocyst yield and quality remained significantly lower than that of oocytes matured in vivo. A significant additive effect of combined IVM treatments was evident as increased COC lactate production and oxygen consumption and enhanced oocyte oxidative metabolism, ATP production, ATP:ADP ratio, and glutathione levels (P < 0.05). Nevertheless, IVM increased reactive oxygen species production, particularly as a consequence of FSH addition, relative to in vivo matured oocytes. In conclusion, improvements in the embryo yield following IVM is associated with increased COC oxygen consumption and oocyte oxidative metabolism, but these remain metabolically and developmentally less competent relative to in vivo derived oocytes.

Quantifying oxygen diffusion in paraffin oil used in oocyte and embryo culture.

Oxygen diffusion through oil is important in the culture of oocytes and embryos. A diffusion coefficient two orders of magnitude smaller than that of oxygen in water has been thought possible, and this has led to concerns of anoxia in cultures. Using an assay for determining the oxygen consumption rate of embryos and oocytes, along with a mathematical model, it is here shown that the oxygen diffusion rate in paraffin oil at 37 degrees C is about two-thirds of that in water at the same temperature. Although not previously recognised for the assay in question, the geometry is such that anoxia does occur for a period of time in excess of 1 hr and, by the completion of the assay, 30-40% of the medium is anoxic. Hence the quantity of oxygen consumed is less than would be consumed in conditions of plentiful oxygen supply. Nevertheless, using a model with a concentration dependent oxygen consumption rate, the oxygen consumption rate can be estimated.

Mathematical modeling of glucose supply toward successful in vitro maturation of Mammalian oocytes.

In vitro maturation-whereby an oocyte is harvested from an ovary just before full maturation, matured in the laboratory, fertilized, and then transplanted back to the uterus-has important benefits over, but is significantly less successful than, traditional in vitro fertilization. Inadequate in vitro nutrient environments are believed to be a prime reason for the low success, but understanding of the in vivo environment, which needs to be better replicated in the laboratory, is still lacking. We here consider mathematical modeling as an aid to increasing that understanding. A general mathematical model suitable for examining the in vivo concentrations of a nutrient in the cumulus-oocyte complex (COC) is presented. We then tailor the model to consider glucose concentration. Experimental data are used to obtain information on glucose uptake in the COC for use in the model. Finally, we solve the model to estimate glucose concentration in the COC. With the information currently available, the model indicates a significant reduction in glucose concentration from the follicular fluid across the cumulus matrix to the oocyte.