Medium- and time-related effects on hypothermic storage of rat testicular cells.

Rageia Elfageih, Ahmed Reda, Kristin Ros Kjartansdottir, Valentina Pampanini, Olle Soder, Jan-Bernd Stukenborg
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Abstract

Objective: Testicular samples obtained for fertility preservation often need to be transported between clinics. This study aimed to mimic this short-term hypothermic storage (4-8 °C) and explore the impact of these conditions and the transport medium composition on prepubertal rat testicular tissue samples.

Methods: Testicular tissue samples obtained from seven days post-partum rats were transferred to six compositionally different basal culture media and a balanced salt solution, which had been kept at 4-8 °C prior to transfer. The samples were preserved for either 12 or 24 hours in these hypothermic conditions. The potential effects of the short-term storage were evaluated by assessing the morphology, measuring the testosterone levels by radioimmunoassay and analysing 96 genes with TaqMan Low-Density Arrays. Summarizing results: Levels of gene expression related to energy, apoptosis and angiogenesis pathways were altered after hypothermic storage for 12 and especially 24 hours. We observed only minor differences in gene expression profiles for germ and testicular somatic cells, and no differences in tissue morphology and testosterone production levels.

Conclusions: Short-term hypothermic storage of testicular tissue with a maximum duration of 24 hours does not affect the overall expression profile of testicular cell-specific genes; however, in a minor way, it affects the expression of specific cellular genes.

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大鼠睾丸细胞低温贮藏的中、时相关效应。
目的:用于保存生育能力的睾丸标本经常需要在诊所之间运输。本研究旨在模拟这种短期低温储存(4-8°C),并探讨这些条件和运输介质组成对青春期前大鼠睾丸组织样品的影响。方法:将产后7 d大鼠睾丸组织样本分别转移至6种成分不同的基础培养基和平衡盐溶液中,转移前将其保存在4-8℃。样品在低温条件下保存12或24小时。通过形态学分析、放射免疫法测定睾丸激素水平和TaqMan低密度阵列分析96个基因,评估短期储存的潜在影响。总结结果:低温保存12小时,尤其是24小时后,能量、凋亡和血管生成通路相关基因表达水平发生改变。我们观察到生殖细胞和睾丸体细胞的基因表达谱只有微小的差异,在组织形态和睾酮产生水平上没有差异。结论:睾丸组织短期低温保存(最长时间24小时)不影响睾丸细胞特异性基因的整体表达谱;然而,它以一种次要的方式影响特定细胞基因的表达。
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