Zhen-Li Zhao , Shao-Hua Hu , Zhi-Sheng Wan , Wei-Zhen Bu , Song-Qiang Chen , Tian-Hong Han , Yi-Qun Lu
{"title":"Effect of icariin on the transformation efficiency of induced pluripotent stem cells into sperm cells in vitro","authors":"Zhen-Li Zhao , Shao-Hua Hu , Zhi-Sheng Wan , Wei-Zhen Bu , Song-Qiang Chen , Tian-Hong Han , Yi-Qun Lu","doi":"10.1016/j.androl.2023.100373","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>To investigate the effect of icariin on the transformation efficiency of germ cell-like cells from mouse induced pluripotent stem cells into sperm cells in vitro.</p></div><div><h3>Methods</h3><p><span>Firstly, mouse induced pluripotent stem cells were induced and cultured to transform into germ cell-like cells, and the primordial germ cell-like cells were identified by Western blot and RT-PCR. Then, different concentrations of icariin (0.1</span> <!-->μg/mL, 1<!--> <!-->μg/mL, 10<!--> <!-->μg/mL and 100<!--> <!-->μg/mL) were added into the culture medium, and the obtained primitive germ cell-like cells were cultured, Western blot and RT-PCR were used to identify the obtained sperm cells, the transformation efficiency was compared.</p></div><div><h3>Results</h3><p>The primordium germ cell-like cells obtained from mouse induced pluripotent stem cells in vitro specially expressed Oct-4 protein, C-kit protein, Mvh mRNA, Fragilis mRNA and Stella mRNA. The sperm cells were specially expressed VASA, SCP3 and γH2AX proteins. RT-PCR showed that the sperm cells were specially expressed Ddx4, Tp2 and Prm1 mRNA. Compared with the control group, the expression level of VASA protein (1.744<!--> <!-->±<!--> <!-->0.283, 2.882<!--> <!-->±<!--> <!-->0.373, 6.489<!--> <!-->±<!--> <!-->0.460), SCP3 protein (2.250<!--> <!-->±<!--> <!-->0.306, 7.058<!--> <!-->±<!--> <!-->0.521, 8.654<!--> <!-->±<!--> <!-->0.804), γH2AX protein (4.304<!--> <!-->±<!--> <!-->0.433, 5.713<!--> <!-->±<!--> <!-->0.339, 9.268<!--> <!-->±<!--> <!-->0.545), Ddx4 mRNA (1.374<!--> <!-->±<!--> <!-->0.145, 2.846<!--> <!-->±<!--> <!-->0.194, 4.021<!--> <!-->±<!--> <!-->0.154), Tp2 mRNA (1.358<!--> <!-->±<!--> <!-->0.130, 3.623<!--> <!-->±<!--> <!-->0.326, 5.811<!--> <!-->±<!--> <!-->0.390) and Prm1 mRNA (1.326<!--> <!-->±<!--> <!-->0.162, 3.487<!--> <!-->±<!--> <!-->0.237, 4.666<!--> <!-->±<!--> <!-->0.307) in 0.1<!--> <!-->μg/mL, 1<!--> <!-->μg/mL, 10<!--> <!-->μg/mL icariin experimental groups were all lower than that of VASA protein (10.560<!--> <!-->±<!--> <!-->0.413), SCP3 protein (13.804<!--> <!-->±<!--> <!-->0.642), γH2AX protein (11.874<!--> <!-->±<!--> <!-->0.464), Ddx4 mRNA (6.4005<!--> <!-->±<!--> <!-->0.361), Tp2 mRNA (7.314<!--> <!-->±<!--> <!-->0.256) and Prm1 mRNA (7.334<!--> <!-->±<!--> <!-->0.390) in 100<!--> <!-->μg/mL icariin experimental group.</p></div><div><h3>Conclusions</h3><p>Icariin can promote the transformation of mouse induced pluripotent stem cells into sperm cells in vitro, and it is concentration-dependent manner in a certain concentration range.</p></div>","PeriodicalId":49129,"journal":{"name":"Revista Internacional De Andrologia","volume":null,"pages":null},"PeriodicalIF":0.8000,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Revista Internacional De Andrologia","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1698031X2300033X","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"ANDROLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Objective
To investigate the effect of icariin on the transformation efficiency of germ cell-like cells from mouse induced pluripotent stem cells into sperm cells in vitro.
Methods
Firstly, mouse induced pluripotent stem cells were induced and cultured to transform into germ cell-like cells, and the primordial germ cell-like cells were identified by Western blot and RT-PCR. Then, different concentrations of icariin (0.1 μg/mL, 1 μg/mL, 10 μg/mL and 100 μg/mL) were added into the culture medium, and the obtained primitive germ cell-like cells were cultured, Western blot and RT-PCR were used to identify the obtained sperm cells, the transformation efficiency was compared.
Results
The primordium germ cell-like cells obtained from mouse induced pluripotent stem cells in vitro specially expressed Oct-4 protein, C-kit protein, Mvh mRNA, Fragilis mRNA and Stella mRNA. The sperm cells were specially expressed VASA, SCP3 and γH2AX proteins. RT-PCR showed that the sperm cells were specially expressed Ddx4, Tp2 and Prm1 mRNA. Compared with the control group, the expression level of VASA protein (1.744 ± 0.283, 2.882 ± 0.373, 6.489 ± 0.460), SCP3 protein (2.250 ± 0.306, 7.058 ± 0.521, 8.654 ± 0.804), γH2AX protein (4.304 ± 0.433, 5.713 ± 0.339, 9.268 ± 0.545), Ddx4 mRNA (1.374 ± 0.145, 2.846 ± 0.194, 4.021 ± 0.154), Tp2 mRNA (1.358 ± 0.130, 3.623 ± 0.326, 5.811 ± 0.390) and Prm1 mRNA (1.326 ± 0.162, 3.487 ± 0.237, 4.666 ± 0.307) in 0.1 μg/mL, 1 μg/mL, 10 μg/mL icariin experimental groups were all lower than that of VASA protein (10.560 ± 0.413), SCP3 protein (13.804 ± 0.642), γH2AX protein (11.874 ± 0.464), Ddx4 mRNA (6.4005 ± 0.361), Tp2 mRNA (7.314 ± 0.256) and Prm1 mRNA (7.334 ± 0.390) in 100 μg/mL icariin experimental group.
Conclusions
Icariin can promote the transformation of mouse induced pluripotent stem cells into sperm cells in vitro, and it is concentration-dependent manner in a certain concentration range.
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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