Mohaqiq Mahdi, Movahedin Mansoureh, Mazaheri Zohreh, Amirjannati Naser
Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. Electronic Address:
Cell J. 2019 Jan;20(4):513-520. doi: 10.22074/cellj.2019.5675. Epub 2018 Aug 1.
In vitro transplantation (IVT) of spermatogonial stem cells (SSCs) is one of the most recent methods in transplantation in recent decades. In this study, IVT and SSCs homing on seminiferous tubules of host testis in organ culture have been studied.
In this experimental study, human SSCs were isolated and their identities were confirmed by tracking their promyelocytic leukemia zinc finger (PLZF) protein. These cells were transplanted to adult azoospermia mouse testes using two methods, namely, IVT and in vivo transplantation as transplantation groups, and testes without transplantation of cells were assigned in the control group. Then histomorphometric, immunohistochemical and molecular studies were done after 2 weeks.
After two weeks, histomorphometric studies revealed that the number of subsided spermatogonial cells (SCs) and the percentage of tubules with subsided SCs in IVT and in vivo groups were significantly more than those in the control group (P<0.05). Immunohistochemical studies in the transplantation groups confirmed that the PLZF protein was expressed in the cells subsided on the seminiferous tubule. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) demonstrated that the PLZF gene expression was only positive in the transplantation groups, but it was not significantly different between the IVT group and the in vivo group (P>0.05).
Testicular tissue culture conditions after SSC transplantation can help these cells subside on the seminiferous tubule basement membrane.
精原干细胞（SSCs）的体外移植（IVT）是近几十年来移植领域最新的方法之一。在本研究中，对器官培养中SSCs在宿主睾丸生精小管上的归巢及IVT进行了研究。
在本实验研究中，分离人SSCs，并通过追踪其早幼粒细胞白血病锌指（PLZF）蛋白来确认其身份。这些细胞采用IVT和体内移植两种方法移植到成年无精子症小鼠睾丸，作为移植组，对照组为未移植细胞的睾丸。然后在2周后进行组织形态计量学、免疫组化和分子研究。
两周后，组织形态计量学研究显示，IVT组和体内移植组中沉降的精原细胞（SCs）数量以及有沉降SCs的小管百分比均显著高于对照组（P<0.05）。移植组的免疫组化研究证实，PLZF蛋白在生精小管上沉降的细胞中表达。定量逆转录聚合酶链反应（qRT-PCR）表明，PLZF基因表达仅在移植组呈阳性，但IVT组和体内移植组之间无显著差异（P>0.05）。
SSC移植后的睾丸组织培养条件可帮助这些细胞沉降在生精小管基底膜上。
