Bilodeau-Goeseels Sylvie
Agriculture and Agri-Food Canada, Lethbridge Research Centre, 5403 1st Ave. South, P.O. Box 3000, Lethbridge, Alta., Canada T1J 4B1.
Theriogenology. 2006 Jul 15;66(2):297-306. doi: 10.1016/j.theriogenology.2005.11.014. Epub 2005 Dec 27.
The influence of the culture medium and energy sources on spontaneous nuclear maturation and inhibition of maturation in bovine cumulus-enclosed oocytes (CEO) was examined. CEO were cultured in Medium 199, minimum essential medium, M16, or synthetic oviduct fluid (SOF), all containing 3 mg/mL bovine serum albumin (BSA), and SOF without BSA, alone or supplemented with hypoxanthine (HYPO, 4 mM) or forskolin (FSK, 100 microM) for 21 h. More CEO remained at the GV stage in M16 compared to other media (P < 0.05). Supplementation with HYPO increased and FSK reduced the percentage of CEO remaining at the GV stage (P < 0.05) only in M16. The effects of energy sources, in the absence or presence of HYPO or FSK, were examined in CEO cultured in M16 salts+PVA. Glucose (0.5 and 5.5 mM), pyruvate (0.32 and 3.2 mM), lactate (3.3 mM) and glutamine (1.3 mM) significantly reduced the percentage of CEO remaining at the GV stage compared to M16 salts alone; only glutamine significantly increased the percentage of CEO at the MII stage compared to M16 salts. In M16 salts+HYPO, glucose (0.5 mM), pyruvate (0.32 mM), lactate (3.3 mM) and glutamine (1.3 mM) significantly reduced the percentage of GV and degenerate oocytes and increased the percentage of CEO at the MI stage. In M16 salts+FSK, the energy sources significantly decreased the percentage of oocytes with condensed chromosomes and increased the percentage of CEO reaching metaphase I. In conclusion, meiotic inhibitors had different effects in different culture media and glucose, pyruvate, lactate and glutamine were stimulatory to nuclear maturation. It was noteworthy that some of the results obtained were contrary to previous findings in mouse oocytes.
研究了培养基和能量来源对牛卵丘-卵母细胞复合体（CEO）自发核成熟及成熟抑制的影响。将CEO分别培养于含3mg/mL牛血清白蛋白（BSA）的M199培养基、基本培养基、M16培养基或合成输卵管液（SOF）以及不含BSA的SOF中，单独培养或添加次黄嘌呤（HYPO，4mM）或福司柯林（FSK，100μM）培养21小时。与其他培养基相比，更多的CEO在M16培养基中停留在生发泡（GV）期（P<0.05）。仅在M16培养基中，添加HYPO可增加而添加FSK可降低停留在GV期的CEO百分比（P<0.05）。在M16盐+聚乙烯醇（PVA）中培养的CEO中，研究了在有无HYPO或FSK存在时能量来源的影响。与单独的M16盐相比，葡萄糖（0.5和5.5mM）、丙酮酸（0.32和3.2mM）、乳酸（3.3mM）和谷氨酰胺（1.3mM）显著降低了停留在GV期的CEO百分比；与M16盐相比，只有谷氨酰胺显著增加了处于减数第二次分裂中期（MII）的CEO百分比。在M16盐+HYPO中，葡萄糖（0.5mM）、丙酮酸（0.32mM）、乳酸（3.3mM）和谷氨酰胺（1.3mM）显著降低了GV期和退化卵母细胞的百分比，并增加了处于减数第一次分裂中期（MI）的CEO百分比。在M16盐+FSK中，能量来源显著降低了染色体凝聚的卵母细胞百分比，并增加了达到中期I的CEO百分比。总之，减数分裂抑制剂在不同培养基中有不同作用，葡萄糖、丙酮酸、乳酸和谷氨酰胺对核成熟有促进作用。值得注意的是，一些获得的结果与先前在小鼠卵母细胞中的发现相反。
