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Enhancing oxygen delivery to ovarian follicles by three different methods markedly improves growth in serum-containing culture medium

J M Connolly, M T Kane, L R Quinlan, A C Hynes
Reproduction, Fertility, and Development 2019 April 12
Invitro ovarian follicle culture systems are routinely used to study folliculogenesis and may provide solutions for infertility. Mouse follicles are typically cultured in standard gas-impermeable culture plates under gas phase oxygen concentrations of 5% or 20% (v/v). There is evidence that these conditions may not provide adequate oxygenation for follicles cultured as non-attached intact units in medium supplemented with serum and high levels of FSH. Three different methods of enhancing follicle oxygenation were investigated in this study: increasing the gas phase oxygen concentration, inverting the culture plates and using gas-permeable culture plates. Follicles cultured under 40% O2 were significantly larger (P<0.01), had increased ovulation rates (P<0.0001) and produced more oestradiol (P<0.05) than follicles cultured under 20% O2. These effects were associated with reduced secretion of vascular endothelial growth factor (P<0.05) and lactate (P<0.05), and reduced expression of hypoxia-related genes. Increasing oxygen delivery with gas-permeable plates or by culture plate inversion also improved follicle growth (P<0.01). An important aspect of enhancing oxygen delivery in this culture system is that it allows development of three-dimensional spherical mouse follicles over 6 days in serum- and FSH-supplemented medium to sizes comparable to invivo-matured follicles (~500μm in diameter). Such follicular development is not possible under hypoxic conditions.


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