Shirzeyli Maryam Hosseinzadeh, Tayyebiazar Ali, Aliakbari Fereshteh, Ghasemi Fahimeh, Eini Fatemeh, Shirzeyli Farhad Hosseinzadeh, Vanaki Elmira, Sobhani Aligholi
Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, I.R. Iran.
Clinical Research Development Unit of Imam Khomeini Hospital, Urmia University of Medical Sciences, Urmia, I.R. Iran.
Res Pharm Sci. 2022 Jan 15;17(2):123-133. doi: 10.4103/1735-5362.335171. eCollection 2022 Apr.
development of functional gametes from pluripotent stem cells is a promising prospect to treat infertility. Mesenchymal stem cells with a high degree of plasticity and less tumorigenicity are a reliable source of stem cells for the generation of gametes. The present study aimed to compare the differentiation potential in the mesenchymal stem cells that are derived from bone marrow (BMDMSCs) and adipose tissue derived mesenchymal stem cells (AD-MSCs) into germ cells in a culture medium containing bone morphogenic protein-4 (BMP-4).
In this study, MSCs were isolated from both bone marrow and adipose tissue of murine samples. To further verify the nature of the harvested stem cells, their multipotency and surface marker were examined. The identified stem cells were cultured in a medium supplemented with 0 and 25 ng/mL of BMP-4 for 4 days. Flow cytometry analysis, immunofluorescence staining, and real RT-PCR were used to assess the expression levels in germ cell-specific biomarkers (Mvh, Dazl, Stra8, and Scp3).
FINDINGS/RESULTS: CD44+, CD45-, CD31-, BMD-MSCs, and AD-MSCs showed to be capable of differentiating to osteo-adipogenic lineages. The flow cytometry, immunofluorescence, and RT-PCR results indicated that early germ cell markers (Mvh and Dazl) were expressed in both types of cells but they were significantly higher in BMD-MSCs than AD-MSCs.
Based on our results, the addition of exogenous BMP4 to the culture medium could differentiate BMD-MSCs and AD-MSCs into primordial germ cells, but it is inadequate to further develop into late germ cells . Moreover, the results revealed that, although AD-MSCs were easier to collect and had faster growth and proliferation rates than BMD-MSCs, the BMD-MSCs were better capable of differentiation into primordial germ cells. They may serve to be considered a more suitable source of MSC for generation of gametes than AD-MSCs.
从多能干细胞发育出功能性配子是治疗不孕症的一个有前景的方向。具有高度可塑性和较低致瘤性的间充质干细胞是用于生成配子的可靠干细胞来源。本研究旨在比较在含有骨形态发生蛋白-4(BMP-4)的培养基中,源自骨髓的间充质干细胞(BMDMSCs)和脂肪组织来源的间充质干细胞(AD-MSCs)向生殖细胞的分化潜能。
在本研究中,从鼠类样本的骨髓和脂肪组织中分离出间充质干细胞。为进一步验证收获的干细胞的性质,检测了它们的多能性和表面标志物。将鉴定出的干细胞在添加有0和25 ng/mL BMP-4的培养基中培养4天。采用流式细胞术分析、免疫荧光染色和实时RT-PCR来评估生殖细胞特异性生物标志物(Mvh、Dazl、Stra8和Scp3)的表达水平。
CD44 +、CD45 -、CD31 -、BMD-MSCs和AD-MSCs显示能够分化为成骨-脂肪生成谱系。流式细胞术、免疫荧光和RT-PCR结果表明,两种类型的细胞中均表达早期生殖细胞标志物(Mvh和Dazl),但在BMD-MSCs中的表达明显高于AD-MSCs。
基于我们的结果,向培养基中添加外源性BMP4可使BMD-MSCs和AD-MSCs分化为原始生殖细胞,但不足以进一步发育为晚期生殖细胞。此外,结果显示,尽管AD-MSCs比BMD-MSCs更容易收集且生长和增殖速度更快,但BMD-MSCs在分化为原始生殖细胞方面能力更强。它们可能被认为是比AD-MSCs更适合用于生成配子的间充质干细胞来源。