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用于人类体外受精的哈姆氏F10培养基的冷冻保存。

Frozen storage of Ham's F10 medium for human in-vitro fertilization.

作者信息

Bernart W, Hauff B, Malez-Kehry W, Kunz G, Leyendecker G

机构信息

Staedtische Frauenklinik, Academic Teaching Hospital, Darmstadt, FRG.

出版信息

Hum Reprod. 1990 Jul;5(5):610-2. doi: 10.1093/oxfordjournals.humrep.a137154.

Abstract

A method of frozen storage of Ham's F10 medium was investigated that provides 'ready-to-use' culture medium for human in-vitro fertilization, without the necessity of readjusting and testing the medium after thawing. Ham's F10 medium, without bicarbonate, was adjusted to 245 mOsm/kg and stored in aliquots of 33 ml at -20 degrees C. Aliquots of 1 ml of a 7.5% (w/v) sodium bicarbonate solution were stored separately at the same temperature. The two components were mixed together after thawing. In the first test series, mouse embryos were cultured in media stored frozen for varying intervals between 2 weeks and 6 months and no difference in the rates of blastocyst formation was detected. Frozen-stored Ham's F10 medium was then used for human IVF in 256 cycles performed within a 16-month period in two different IVF centres. The pregnancy rates were evaluated and correlated with the duration of the frozen storage (between 1 week and 3 months) and compared to the outcome of 24 cases in which non-frozen medium was used. There was no significant difference in the pregnancy rates in the different groups (19% with non-frozen medium and between 21 and 33% with frozen-stored medium). Thus it was shown that there is no loss of quality of the frozen-stored media within the tested period of 3 months. The prolonged storage interval offers the possibility of extended quality tests and cross-tests between different IVF laboratories.

摘要

研究了一种Ham's F10培养基的冷冻保存方法,该方法可为人类体外受精提供“即用型”培养基,解冻后无需重新调整和测试培养基。将不含碳酸氢盐的Ham's F10培养基调整至245 mOsm/kg,分装成33 ml的 aliquots,于-20℃保存。将1 ml 7.5%(w/v)的碳酸氢钠溶液分装,在相同温度下单独保存。解冻后将两种成分混合在一起。在第一个测试系列中,将小鼠胚胎培养在冷冻保存不同时间间隔(2周和6个月之间)的培养基中,未检测到囊胚形成率有差异。然后,在两个不同的体外受精中心,在16个月内进行的256个周期的人类体外受精中使用了冷冻保存的Ham's F10培养基。评估了妊娠率,并将其与冷冻保存时间(1周和3个月之间)相关联,并与24例使用非冷冻培养基的结果进行了比较。不同组的妊娠率没有显著差异(非冷冻培养基为19%,冷冻保存培养基为21%至33%)。因此表明,在3个月的测试期内,冷冻保存的培养基质量没有损失。延长的保存间隔为不同体外受精实验室之间进行扩展的质量测试和交叉测试提供了可能性。

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