Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran.
J Assist Reprod Genet. 2022 Jan;39(1):153-163. doi: 10.1007/s10815-021-02309-8. Epub 2021 Sep 14.
The generation of germ cells from mesenchymal stromal cells (MSCs) provides a valuable in vitro platform for infertility modeling. The establishment of these cells is a new approach for assisted reproductive technology (ART) to help infertile patients who lack functional gametes.
Human adipose-derived MSCs were isolated and then characterized for multipotency by flow cytometry, differentiation capacity, and cytogenetic assays. These cells were used in a male germ cell differentiation study. The expression of male germ cell markers was evaluated at day 21 of differentiation using an immunofluorescence assay, flow cytometry, and RT-qPCR. Undifferentiated MSCs were used for transplantation in busulfan-induced azoospermic mice.
In this study, MSCs were successfully isolated from human adipose tissues which were positive for cell markers such as CD90, CD105, CD73, and CD29 but negative for CD34 and CD45. The results of flow cytometry, immunocytochemistry, and RT-qPCR analysis at day 21 of differentiation showed that the undifferentiated adipose-derived MSCs are able to differentiate into male germ cells. Additionally, transplantation of undifferentiated MSCs in busulfan-induced azoospermic mice caused spermatogenesis recovery in the majority of seminiferous tubules.
In this study, we showed that differentiation of human adipose-derived MSCs into male germ cells is a useful tool for in vitro study of human germ cell development. Our results demonstrated that cell therapy with adipose-derived MSCs could help the repair of pathological changes in testicular seminiferous tubules. Therefore, it may have a clinical application for the treatment of azoospermia in infertile patients.
从间充质基质细胞（MSCs）中生成生殖细胞为不孕建模提供了有价值的体外平台。这些细胞的建立是辅助生殖技术（ART）的一种新方法，可以帮助缺乏功能性配子的不孕患者。
分离人脂肪来源的 MSCs，然后通过流式细胞术、分化能力和细胞遗传学检测对其多能性进行鉴定。这些细胞用于男性生殖细胞分化研究。使用免疫荧光、流式细胞术和 RT-qPCR 检测分化第 21 天的男性生殖细胞标志物表达情况。未分化的 MSCs 用于移植到白消安诱导的无精子症小鼠中。
在这项研究中，成功地从人脂肪组织中分离出 MSCs，这些细胞对细胞标志物呈阳性，如 CD90、CD105、CD73 和 CD29，但对 CD34 和 CD45 呈阴性。分化第 21 天的流式细胞术、免疫细胞化学和 RT-qPCR 分析结果表明，未分化的脂肪来源 MSCs 能够分化为男性生殖细胞。此外，将未分化的 MSCs 移植到白消安诱导的无精子症小鼠中，可使大多数曲细精管中的精子发生恢复。
在这项研究中，我们表明人脂肪来源 MSCs 分化为男性生殖细胞是体外研究人类生殖细胞发育的有用工具。我们的结果表明，脂肪来源的 MSCs 细胞治疗可以帮助修复睾丸曲细精管的病变。因此，它可能在治疗不孕患者的无精子症方面具有临床应用价值。
