a Department of Anatomical Sciences, Faculty of Medical Science , Tarbiat Modares University , Tehran , Iran.
b Department of Anatomical Sciences, Basic Medical Research Center , Histogenotech Company , Tehran , Iran.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):1585-1594. doi: 10.1080/21691401.2019.1594854.
This study aimed to induce spermatogenesis in azoospermic testis through induced pluripotent stem cells (iPSCs) derived spermatogonial stem cell-like cells (SSCLCs) after iPSCs in vivo and in vitro transplantation and three-dimensional organ culture. DiI-labelled mouse iPSCs were transplanted to azoospermic testis mouse model (pretreated by busulfan 40 mg/kg). This study was designed based on two experimental groups. In experimental group 1(in vivo) labelled iPSCs were transplanted to azoospermic host testis. In experimental group 2 (in vitro) after cell transplantation, fragments of host testes were set as 3D organ culture and testis without cells transplantation served as the control group by the same method. The samples were evaluated by tracing DiI, cell homing, immunohistochemistry, and quantitative RT PCR assays. 2 weeks after iPSCs transplantation, the molecular assessment showed that Plzf, Thy1, Vasa and Gfra1 expression were increased significantly (p ≤ .05) in host testis and labelled iPSCs co-localized by the Plzf and Thy1 markers expression in the base of seminiferous tubules. These findings suggest the ability of iPSCs to achieve homing in the testis niche and indicate the critical inductive role of microenvironment signals in the differentiation of iPSCs to spermatogonial stem cell-like cells.
本研究旨在通过诱导多能干细胞(iPSCs)衍生的精原干细胞样细胞(SSCLCs)在体内和体外移植以及三维器官培养后,诱导无精子症睾丸中的精子发生。用 DiI 标记的小鼠 iPSCs 移植到无精子症睾丸小鼠模型(用 40mg/kg 白消安预处理)。本研究基于两个实验组进行设计。在实验组 1(体内)中,将标记的 iPSCs 移植到无精子症宿主睾丸中。在实验组 2(体外)中,在细胞移植后,将宿主睾丸的碎片设置为 3D 器官培养,并用相同方法将没有细胞移植的睾丸作为对照组。通过追踪 DiI、细胞归巢、免疫组织化学和定量 RT-PCR 分析评估样本。在 iPSCs 移植后 2 周,分子评估显示,宿主睾丸和 Plzf、Thy1 标志物表达阳性的标记 iPSCs 中 Plzf、Thy1、Vasa 和 Gfra1 的表达显著增加(p≤.05),这些发现表明 iPSCs 具有归巢到睾丸龛位的能力,并表明微环境信号在 iPSCs 向精原干细胞样细胞分化中的关键诱导作用。