Investigation of Signals and Transcription Factors for The Generation of Female Germ-Like Cells
Objective: Primordial germ cell (PGCs) lines are a source of a highly specialized type of cells, characteristically oocytes,during female germline development in vivo. The oocyte growth begins in the transition from the primary follicle. It isassociated with dynamic changes in gene expression, but t...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Royan Institute (ACECR), Tehran
2022-08-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | https://www.celljournal.org/article_253358_34cb6f5fe59acdf36bb65e6a7bafab7f.pdf |
| _version_ | 1811183378261606400 |
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| author | Saman Ebrahimi Alireza Shams Parvaneh Maghami Azadeh Hekmat |
| author_facet | Saman Ebrahimi Alireza Shams Parvaneh Maghami Azadeh Hekmat |
| author_sort | Saman Ebrahimi |
| collection | DOAJ |
| description | Objective: Primordial germ cell (PGCs) lines are a source of a highly specialized type of cells, characteristically oocytes,during female germline development in vivo. The oocyte growth begins in the transition from the primary follicle. It isassociated with dynamic changes in gene expression, but the gene-regulating signals and transcription factors that control oocyte growth remain unknown. We aim to investigate the differentiation potential of mouse bone marrow mesenchymal stem cells (mMSCs) into female germ-like cells by testing several signals and transcription factors.Materials and Methods: In this experimental study, mMSCs were extracted from mice femur bone using the flushingtechnique. The cluster-differentiation (CD) of superficial mesenchymal markers was determined with flow cytometric analysis. We applied a set of transcription factors including retinoic acid (RA), titanium nanotubes (TNTs), and fibrin such as TNT-coated fibrin (F+TNT) formation and (RA+F+TNT) induction, and investigated the changes in gene, MVH/ DDX4, expression and functional screening using an in vitro mouse oocyte development condition. Germ cell markers expression, (MVH / DDX4), was analyzed with Immunocytochemistry staining, quantitative transcription-polymerase chain reaction (RT-qPCR) analysis, and Western blots.Results: The expression of CD was confirmed by flow cytometry. The phase determination of the TNTs and F+TNT were confirmed using x-ray diffraction (XRD) and scanning electron microscope (SEM), respectively. Remarkably, applying these transcription factors quickly induced pluripotent stem cells into oocyte-like cells that were sufficient to generate female germlike cells, growth, and maturation from mMSCs differentiation. These transcription factors formed oocyte-like cells specification of stem cells, epigenetic reprogramming, or meiosis and indicate that oocyte meiosis initiation and oocyte growth are not separable from the previous epigenetic reprogramming in stem cells in vitro.Conclusion: Results suggested several transcription factors may apply for arranging oocyte-like cell growth and supplies an alternative source of in vitro maturation (IVM). |
| first_indexed | 2024-04-11T09:46:41Z |
| format | Article |
| id | doaj.art-b1e4f34f6f7440458d88c04f7763588a |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-04-11T09:46:41Z |
| publishDate | 2022-08-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-b1e4f34f6f7440458d88c04f7763588a2022-12-22T04:30:58ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142022-08-0124845846410.22074/cellj.2022.8303253358Investigation of Signals and Transcription Factors for The Generation of Female Germ-Like CellsSaman Ebrahimi0Alireza Shams1Parvaneh Maghami2Azadeh Hekmat3Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, IranDepartment of Anatomy, School of Medicine, Alborz University of Medical Sciences, Karaj, IranDepartment of Biology, Science and Research Branch, Islamic Azad University, Tehran, IranDepartment of Biology, Science and Research Branch, Islamic Azad University, Tehran, IranObjective: Primordial germ cell (PGCs) lines are a source of a highly specialized type of cells, characteristically oocytes,during female germline development in vivo. The oocyte growth begins in the transition from the primary follicle. It isassociated with dynamic changes in gene expression, but the gene-regulating signals and transcription factors that control oocyte growth remain unknown. We aim to investigate the differentiation potential of mouse bone marrow mesenchymal stem cells (mMSCs) into female germ-like cells by testing several signals and transcription factors.Materials and Methods: In this experimental study, mMSCs were extracted from mice femur bone using the flushingtechnique. The cluster-differentiation (CD) of superficial mesenchymal markers was determined with flow cytometric analysis. We applied a set of transcription factors including retinoic acid (RA), titanium nanotubes (TNTs), and fibrin such as TNT-coated fibrin (F+TNT) formation and (RA+F+TNT) induction, and investigated the changes in gene, MVH/ DDX4, expression and functional screening using an in vitro mouse oocyte development condition. Germ cell markers expression, (MVH / DDX4), was analyzed with Immunocytochemistry staining, quantitative transcription-polymerase chain reaction (RT-qPCR) analysis, and Western blots.Results: The expression of CD was confirmed by flow cytometry. The phase determination of the TNTs and F+TNT were confirmed using x-ray diffraction (XRD) and scanning electron microscope (SEM), respectively. Remarkably, applying these transcription factors quickly induced pluripotent stem cells into oocyte-like cells that were sufficient to generate female germlike cells, growth, and maturation from mMSCs differentiation. These transcription factors formed oocyte-like cells specification of stem cells, epigenetic reprogramming, or meiosis and indicate that oocyte meiosis initiation and oocyte growth are not separable from the previous epigenetic reprogramming in stem cells in vitro.Conclusion: Results suggested several transcription factors may apply for arranging oocyte-like cell growth and supplies an alternative source of in vitro maturation (IVM).https://www.celljournal.org/article_253358_34cb6f5fe59acdf36bb65e6a7bafab7f.pdfcell differentiationgerm cellstranscription factors |
| spellingShingle | Saman Ebrahimi Alireza Shams Parvaneh Maghami Azadeh Hekmat Investigation of Signals and Transcription Factors for The Generation of Female Germ-Like Cells Cell Journal cell differentiation germ cells transcription factors |
| title | Investigation of Signals and Transcription Factors for The Generation of Female Germ-Like Cells |
| title_full | Investigation of Signals and Transcription Factors for The Generation of Female Germ-Like Cells |
| title_fullStr | Investigation of Signals and Transcription Factors for The Generation of Female Germ-Like Cells |
| title_full_unstemmed | Investigation of Signals and Transcription Factors for The Generation of Female Germ-Like Cells |
| title_short | Investigation of Signals and Transcription Factors for The Generation of Female Germ-Like Cells |
| title_sort | investigation of signals and transcription factors for the generation of female germ like cells |
| topic | cell differentiation germ cells transcription factors |
| url | https://www.celljournal.org/article_253358_34cb6f5fe59acdf36bb65e6a7bafab7f.pdf |
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