Characterization of embryonic stem-like cells derived from mouse spermatogonial stem cells following low-intensity ultrasound treatment

IF 0.8 4区医学 Q4 ANDROLOGY

Revista Internacional De Andrologia Pub Date : 2021-10-01 DOI:10.1016/j.androl.2020.05.010

Arian Azimi , Mahdi Mohaqiq , Mansoureh Movahedin , Zohreh Mazaheri

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引用次数: 1

Abstract

Objective

Spermatogonial stem cells (SSCs) are able to form embryonic stem-like cells (ES-like cells) and embryonic bodies (EBs). Low-intensity ultrasound stimulation (LIUS) has positive effects on the growth and differentiation of the different cells. In this study, we tried to investigate the effects of LIUS on SSC differentiation to ES-like cells.

Materials and methods

SSCs were isolated from neonatal mice and their identification was confirmed by tracking of PLZF, Oct-4, and C-Kit proteins. The SSCs and Sertoli cells were co-cultured in DMEM/F12 supplemented with 15% FBS and LIF. SSCs stimulated by LIUS with 200 mW/CM² intensity. Characterization of obtained ES-like cells was confirmed with Sox2, Oct-4, and SSEA-1 immunofluorescence staining. Also, real-time PCR was performed to analyse the expression of c-Myc and Nanog genes in ES-Like Cells and Stra8, Piwil2 and Plzf genes in SSCs after 21 days of the in vitro culture.

Results

Our results showed c-Kit, PLZF and Oct-4 proteins were expressed positively in SSCs and Sox2, Oct-4, SSEA-1 in the ES-like cells by immunocytochemistry. The results of flow cytometry showed a significant increase in expression of c-Myc and Nanog in ES-like cells compared to SSCs (p < .05), whereas the Stra8, Piwil2, and Plzf became down-regulated during 21 days of culture. ES-like markers cell SSEA-1, Sox2 and Oct-4 were increased in the LIUS group compared to the control group (p < .05).

Conclusion

The results indicated that ES-like cells with pluripotency characteristics were derived from SSCs.

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低强度超声处理后小鼠精原干细胞衍生的胚胎干细胞样细胞的特性

目的永原干细胞(SSCs)具有形成胚胎干细胞样细胞(ES-like cells)和胚胎体(EBs)的能力。低强度超声刺激(LIUS)对不同细胞的生长和分化有积极作用。在本研究中，我们试图探讨LIUS对SSC向es样细胞分化的影响。材料和方法从新生小鼠中分离ssscs，通过PLZF、Oct-4和C-Kit蛋白的追踪鉴定ssscs。SSCs和Sertoli细胞在添加15% FBS和LIF的DMEM/F12中共培养。用200 mW/CM2强度的LIUS刺激ssc。用Sox2、Oct-4和SSEA-1免疫荧光染色证实获得的es样细胞的特征。体外培养21 d后，采用实时荧光定量PCR方法分析ES-Like细胞中c-Myc和Nanog基因的表达情况，以及SSCs中Stra8、Piwil2和Plzf基因的表达情况。结果免疫细胞化学结果显示，sc中c-Kit、PLZF和Oct-4蛋白阳性表达，es样细胞中Sox2、Oct-4、SSEA-1蛋白阳性表达。流式细胞术结果显示，与ssc相比，es样细胞中c-Myc和Nanog的表达显著增加(p <.05)，而Stra8、Piwil2和Plzf在培养21天后下调。与对照组相比，LIUS组es样标记细胞SSEA-1、Sox2和Oct-4升高(p <. 05)。结论胚胎干细胞可获得具有多能性的es样细胞。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Revista Internacional De Andrologia ANDROLOGY-

CiteScore

1.60

自引率

12.50%

发文量

审稿时长

>12 weeks

期刊介绍： Revista Internacional de Andrología es la revista oficial de la Asociación Española de Andrología, Medicina Sexual y Reproductiva (ASESA), la Sociedade Portuguesa de Ardrologia, la Sociedad Argentina de Andrología (SAA), la Asociación Iberoamericana de Sociedades de Andrología (ANDRO), y la Federación Española de Sociedades de Sexología. La revista publicada trimestralmente es revisada por pares y es líder en el la especialidad y en español y portugués. Recientemente también publica artículos en inglés. El objetivo de la revista es principalmente la promoción del conocimiento y la educación médica continua, con un enfoque especial en el público español y latinoamericano, a través de la publicación de las contribuciones importantes de la investigación en el campo. Todos los miembros de las sociedades antes mencionadas reciben la revista y otros suscriptores individuales e institucionales de España, Portugal y América Latina.

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