{"title":"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","authors":"Arian Azimi , Mahdi Mohaqiq , Mansoureh Movahedin , Zohreh Mazaheri","doi":"10.1016/j.androl.2020.05.010","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>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.</p></div><div><h3>Materials and methods</h3><p>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/CM<sup>2</sup> 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.</p></div><div><h3>Results</h3><p>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 (<em>p</em> <!--><<!--> <!-->.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 (<em>p</em> <!--><<!--> <!-->.05).</p></div><div><h3>Conclusion</h3><p>The results indicated that ES-like cells with pluripotency characteristics were derived from SSCs.</p></div>","PeriodicalId":49129,"journal":{"name":"Revista Internacional De Andrologia","volume":null,"pages":null},"PeriodicalIF":0.8000,"publicationDate":"2021-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Characterization of embryonic stem-like cells derived from mouse spermatogonial stem cells following low-intensity ultrasound treatment\",\"authors\":\"Arian Azimi , Mahdi Mohaqiq , Mansoureh Movahedin , Zohreh Mazaheri\",\"doi\":\"10.1016/j.androl.2020.05.010\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><p>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.</p></div><div><h3>Materials and methods</h3><p>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/CM<sup>2</sup> 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.</p></div><div><h3>Results</h3><p>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 (<em>p</em> <!--><<!--> <!-->.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 (<em>p</em> <!--><<!--> <!-->.05).</p></div><div><h3>Conclusion</h3><p>The results indicated that ES-like cells with pluripotency characteristics were derived from SSCs.</p></div>\",\"PeriodicalId\":49129,\"journal\":{\"name\":\"Revista Internacional De Andrologia\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.8000,\"publicationDate\":\"2021-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Revista Internacional De Andrologia\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1698031X20300765\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"ANDROLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Revista Internacional De Andrologia","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1698031X20300765","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"ANDROLOGY","Score":null,"Total":0}
Characterization of embryonic stem-like cells derived from mouse spermatogonial stem cells following low-intensity ultrasound treatment
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.
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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