偵測pyruvate之消耗與lactate之產生可評估培養液之品質
http://humrep.oxfordjournals.org/content/17/11/2950.full
Culture environment modulates maturation and metabolism of human oocytes
+Author Affiliations
- Received January 8, 2002.
- Revision received March 19, 2002.
- Accepted June 12, 2002.
Abstract
BACKGROUND: The clinical use of oocytes matured in vitro for IVF is increasing, but little is known about the effect of culture conditions on oocyte maturation. METHODS: Denuded immature oocytes identified following superovulation prior to ICSI were individually matured in one of two commercial media: tissue culture medium (TCM) 199 or modified Eagle medium with Earle’s modified salts (MEME). During maturation, depletion of pyruvate and accumulation of lactate in the culture medium were non-invasively measured. RESULTS: Maturing oocytes took up pyruvate (20–30 pmol/oocyte/h) and produced lactate (2–10 pmol/oocyte/h). Oocytes matured faster in MEME, with significantly more oocytes reaching metaphase II by 24 h after oocyte retrieval compared with TCM 199 (P = 0.03). The oocytes that matured more quickly in MEME had significantly lower lactate production than oocytes that matured more slowly (P = 0.02). In TCM 199, pyruvate uptake by rapidly maturing oocytes was lower than by slowly maturing oocytes (P = 0.05). During the second incubation, from 24 to 48 h post-oocyte retrieval, pyruvate uptake in MEME was 30% lower than in TCM 199 (P = 0.007). Pyruvate uptake and lactate production differed depending on the stage of nuclear maturation: pyruvate uptake and lactate production were greater during germinal vesicle breakdown than during polar body extrusion in MEME (P < 0.05). CONCLUSIONS: We have shown that: (i) pyruvate is a major energy source during oocyte maturation; (ii) the composition of the culture medium can affect the rate of maturation; and (iii) the culture medium and stage of nuclear maturation can affect pyruvate uptake and lactate production.
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