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首页> 外文期刊>Cell and Tissue Research >Effect of cryopreservation on human granulosa cell viability and responsiveness to gonadotropin
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Effect of cryopreservation on human granulosa cell viability and responsiveness to gonadotropin

机译：冷冻保存对人颗粒细胞活力和对促性腺激素的反应性的影响

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In human, the use of freshly recovered granulosa cells for experiments remains difficult. Because of the single use of human cells, the experiments cannot be repeated, and no additional conditions can be tested afterwards with the cells of the same patient. Therefore, granulosa cell cryopreservation could be a good alternative to keep part of these cells for later controls or experiments. The aim of this study is to compare the responsiveness to FSH of fresh and frozen-thawed human primary granulosa-lutein cells (hGLC) and determine if cryopreserved granulosa cells can be used in place of fresh cells. Two cryopreservation methods were also compared: a conventional versus a simplified freezing method. This experimental study was undertaken at Igyxos S.A., Nouzilly, France. Seventy women undergoing oocyte retrieval at the IVF Unit from Bretonneau University Hospital (Tours, France) were recruited in 2016. Fresh and frozen-thawed hGLC were cultured for 7 days and then stimulated by r-FSH for 48 h. To assess r-FSH efficacy and potency, extracellular cAMP accumulated in the supernatant for each stimulation point was measured. We demonstrated that hGLC remain responsive to FSH stimulation after freezing-thawing and 7 days of pre-culture. They are able to secrete cAMP with a similar EC50 value as fresh hGLC, but FSH efficacy is lowered. As our study did not show any significant difference between the two freezing methods concerning the sensitivity of hGLC to FSH, hGLC could be cryopreserved with the simplified freezing method without taking up too much time for IVF laboratories.
机译：在人类中，使用新恢复的颗粒细胞进行实验仍然困难。由于单一使用人体细胞，实验不能重复，并且之后没有额外的条件与同一患者的细胞进行测试。因此，颗粒细胞冷冻保存可以是使这些细胞的一部分用于后续对照或实验的替代方案。本研究的目的是比较对新鲜和冷冻的人原粒颗粒细胞（HGLC）的FSH的反应能力，并确定是否可以使用冷冻保存的颗粒细胞代替新鲜细胞。还比较了两个冷冻保存方法：常规与简化的冷冻方法。该实验研究是在法国的Igyxos S.A.A.A.A.，Nouzilly的实验研究。 2016年招募了七十名妇女在福尔松大学医院（法国之旅）的IVF单位的卵母线检索。新鲜和冷冻的Hglc被培养7天，然后通过R-FSH刺激48小时。为了评估R-FSH功效和效力，测量每个刺激点上的上清液中积聚的细胞外阵马。我们证明HGLC在冷冻解冻和7天预培养后对FSH刺激保持敏感。他们能够分泌与类似的EC50值为新鲜HGLC的阵营，但是FSH功效降低。由于我们的研究没有显示出关于HGLC对FSH的敏感性的冻结方法之间的任何显着差异，可以用简化的冻结方法冷冻保存HGLC，而不会为IVF实验室占用太多时间。

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