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对单个人类卵母细胞和植入前胚胎的葡萄糖和丙酮酸摄取进行无创测量。

Non-invasive measurement of glucose and pyruvate uptake by individual human oocytes and preimplantation embryos.

作者信息

Hardy K, Hooper M A, Handyside A H, Rutherford A J, Winston R M, Leese H J

机构信息

Institute of Obstetrics and Gynaecology, Hammersmith Hospital, London, UK.

出版信息

Hum Reprod. 1989 Feb;4(2):188-91. doi: 10.1093/oxfordjournals.humrep.a136869.

Abstract

Pyruvate and glucose uptake by 73 individual human oocytes and preimplantation embryos was measured non-invasively, using an ultramicrofluorescence assay to analyse changes in substrate levels in microdroplets of culture medium. The uptake of both substrates was measured over successive daily incubations between days 1 (unfertilized oocytes) or 2 ('spare' embryos which were not transferred) and day 6 (day 0 = day of insemination). Under these conditions, 58% (25/43) of fertilized embryos with two pronuclei on day 1 developed to the blastocyst stage by day 6. The pyruvate uptake of these embryos increased from approximately 28 to a maximum of 40 pmol/embryo/h between days 2.5 and 4.5. Similarly, glucose uptake increased from approximately 8 to 14 pmol/embryo/h between days 2.5 and 4.5, but then increased further to 24 pmol/embryo/h on day 5 at the blastocyst stage. [corrected] The pyruvate uptake of fertilized embryos which arrested at cleavage stages was significantly lower than for those which developed to the blastocyst stage. Polyspermic and parthenogenetic embryos, and unfertilized oocytes also had lower pyruvate uptakes at later stages. The glucose uptake of unfertilized oocytes and abnormal embryos never reached the level of fertilized embryos at the blastocyst stage on day 5.5. Non-invasive measurement of pyruvate uptake before embryo transfer may provide a valuable functional criterion for the selection of viable embryos capable of developing to the blastocyst stage.

摘要

采用超微量荧光分析法,通过分析培养基微滴中底物水平的变化,对73个单个的人类卵母细胞和植入前胚胎的丙酮酸和葡萄糖摄取进行了非侵入性测量。在第1天(未受精的卵母细胞)或第2天(未移植的“备用”胚胎)至第6天(第0天=授精日)之间,连续每日培养期间测量两种底物的摄取情况。在这些条件下,第1天有两个原核的受精胚胎中,58%(25/43)在第6天发育到囊胚阶段。这些胚胎的丙酮酸摄取量在第2.5天至4.5天之间从约28皮摩尔/胚胎/小时增加到最高40皮摩尔/胚胎/小时。同样,葡萄糖摄取量在第2.5天至4.5天之间从约8皮摩尔/胚胎/小时增加到14皮摩尔/胚胎/小时,但在第5天囊胚阶段进一步增加到24皮摩尔/胚胎/小时。[已修正] 在卵裂阶段停滞的受精胚胎的丙酮酸摄取量显著低于发育到囊胚阶段的胚胎。多精受精和孤雌生殖胚胎以及未受精的卵母细胞在后期的丙酮酸摄取量也较低。未受精的卵母细胞和异常胚胎的葡萄糖摄取量在第5.5天囊胚阶段从未达到受精胚胎的水平。胚胎移植前丙酮酸摄取的非侵入性测量可能为选择能够发育到囊胚阶段的 viable 胚胎提供有价值的功能标准。

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