在实验性无精子症小鼠模型中，用生长因子培养的脂肪组织来源间充质干细胞移植后的精子发生恢复潜力。
Spermatogenesis Recovery Potentials after Transplantation of Adipose Tissue-Derived Mesenchymal Stem Cells Cultured with Growth Factors in Experimental Azoospermic Mouse Models.
作者信息
Eliyasi Dashtaki Masoumeh, Hemadi Masoud, Saki Ghasem, Mohammadiasl Javad, Khodadadi Ali
机构信息
Cellular and Molecular Research Center, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Physiology Research Center, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran. Electronic Address:
出版信息
Cell J. 2020 Jan;21(4):401-409. doi: 10.22074/cellj.2020.6055. Epub 2019 Jul 29.
OBJECTIVE
Approximately 1% of the male population suffers from obstructive or non-obstructive azoospermia. Previous in vitro studies have successfully differentiated mesenchymal stem cells (MSCs) into germ cells. Because of immunemodulating features, safety, and simple isolation, adipose tissue-derived MSCs (AT-MSCs) are good candidates for such studies. However, low availability is the main limitation in using these cells. Different growth factors have been investigated to overcome this issue. In the present study, we aimed to comparatively assess the performance of AT-MSCs cultured under the presence or absence of three different growth factors, epidermal growth factor (EGF), leukemia inhibitory factor (LIF) and glial cell line-derived neurotrophic factor (GDNF), following transplantation in testicular torsion-detorsion mice.
MATERIALS AND METHODS
This was an experimental study in which AT-MSCs were first isolated from male Naval Medical Research Institute (NMRI) mice. Then, the mice underwent testicular torsion-detorsion surgery and received bromodeoxyuridine (BrdU)-labeled AT-MSCs into the lumen of seminiferous tubules. The transplanted cells had been cultured in different conditioned media, containing the three growth factors and without them. The expression of germ cell-specific markers was evaluated with real-time polymerase chain reaction (PCR) and western-blot. Moreover, immunohistochemical staining was used to trace the labeled cells.
RESULTS
The number of transplanted AT-MSCs resided in the basement membrane of seminiferous tubules significantly increased after 8 weeks. The expression levels of and genes in the transplanted testicles by AT-MSCs cultured in the growth factors-supplemented medium was greater than those in the control group (P<0.001 and P<0.05, respectively). The expression levels of the and genes did not significantly differ from the control group.
CONCLUSION
Our findings showed that the use of EGF, LIF and GDNF to culture AT-MSCs can be very helpful in terms of MSC survival and localization.
目的
约1%的男性人口患有梗阻性或非梗阻性无精子症。先前的体外研究已成功将间充质干细胞（MSCs）分化为生殖细胞。由于脂肪组织来源的间充质干细胞（AT-MSCs）具有免疫调节特性、安全性高且分离简单，因此是此类研究的良好候选者。然而，细胞可用性低是使用这些细胞的主要限制因素。人们已经研究了不同的生长因子来克服这个问题。在本研究中，我们旨在比较评估在三种不同生长因子，即表皮生长因子（EGF）、白血病抑制因子（LIF）和胶质细胞系源性神经营养因子（GDNF）存在或不存在的情况下培养的AT-MSCs，在睾丸扭转-复位小鼠体内移植后的表现。
材料与方法
这是一项实验研究，首先从雄性海军医学研究所（NMRI）小鼠中分离出AT-MSCs。然后，对小鼠进行睾丸扭转-复位手术，并将溴脱氧尿苷（BrdU）标记的AT-MSCs注入生精小管腔内。移植细胞在含有这三种生长因子和不含生长因子的不同条件培养基中培养。用实时聚合酶链反应（PCR）和蛋白质免疫印迹法评估生殖细胞特异性标志物的表达情况。此外，免疫组织化学染色用于追踪标记细胞。
结果
8周后，位于生精小管基底膜的移植AT-MSCs数量显著增加。在添加生长因子的培养基中培养的AT-MSCs移植到睾丸后， 和 基因的表达水平高于对照组（分别为P<0.001和P<0.05）。 和 基因的表达水平与对照组无显著差异。
结论
我们的研究结果表明，使用EGF、LIF和GDNF培养AT-MSCs在MSC存活和定位方面非常有帮助。
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