Department of Anatomical Sciences, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
Department of Anatomical Sciences, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran; Physiology Research Centre, Kerman University of Medical Sciences, Kerman, Iran; Neuroscience Research Centre, Kerman University of Medical Sciences, Kerman, Iran.
Cytotherapy. 2014 Feb;16(2):203-12. doi: 10.1016/j.jcyt.2013.10.015.
Vitrification as an advanced cryopreservation method is recommended for cell storage toward future applications. The purpose of this report was to appraise whether gametogenic potential of these cells is altered by vitrification.
A two steps method was applied for hUCM cells vitrification. An n-hUCM group of hUCM cells served as control. In order to differentiation of hUCM cells into male germ cells, the cells were induced by retinoic acid, testosterone and testicular-cell-conditioned medium. To evaluate induced hUCM cells toward germ cells, we used immunocytochemistry and karyotyping methods.
v-hUCM cells similar to n-hUCM cells formed flat cells after gametogenic induction, and showed protein expression of germ-cell-specific markers DAZL, VASA (DDX4) and SCP3. Karyotyping pattern remained unchanged in the either groups.
The analysis of these results demonstrates that vitrification does not alter differentiation potential of hUCMs to male germ like cells. These results may set an in vitro pattern to study germ-cell formation from hUCM cells and also as a potential source of sperms for male infertility.
玻璃化作为一种先进的冷冻保存方法，推荐用于细胞储存，以满足未来的应用需求。本报告旨在评估这些细胞的生殖潜能是否因玻璃化而改变。
采用两步法对 hUCM 细胞进行玻璃化处理。n-hUCM 组的 hUCM 细胞作为对照。为了将 hUCM 细胞分化为雄性生殖细胞，用维甲酸、睾酮和睾丸细胞条件培养基诱导细胞。为了评估诱导的 hUCM 细胞向生殖细胞的分化，我们使用免疫细胞化学和核型分析方法。
与 n-hUCM 细胞类似，v-hUCM 细胞在生殖诱导后形成扁平细胞，并显示出生殖细胞特异性标志物 DAZL、VASA（DDX4）和 SCP3 的蛋白表达。两组的核型模式均未改变。
这些结果的分析表明，玻璃化不会改变 hUCM 向雄性生殖样细胞分化的潜能。这些结果可能为从 hUCM 细胞中研究生殖细胞形成提供一种体外模式，也可能成为男性不育症精子的潜在来源。
