Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cells
Abstract Background Embryonic Sertoli cells (eSCs) play an important role in sex determination and in male gonad development which makes them a very useful cell type for therapeutic applications. However, the deriving mechanism of Sertoli cells has been unclear and challenging to create a large numb...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-03-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13287-019-1180-6 |
| _version_ | 1828803074882273280 |
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| author | Chenze Xu Ali Mohsin Yanxia Luo Lili Xie Yan Peng Qizheng Wang Haifeng Hang Yingping Zhuang Meijin Guo |
| author_facet | Chenze Xu Ali Mohsin Yanxia Luo Lili Xie Yan Peng Qizheng Wang Haifeng Hang Yingping Zhuang Meijin Guo |
| author_sort | Chenze Xu |
| collection | DOAJ |
| description | Abstract Background Embryonic Sertoli cells (eSCs) play an important role in sex determination and in male gonad development which makes them a very useful cell type for therapeutic applications. However, the deriving mechanism of Sertoli cells has been unclear and challenging to create a large number of quality eSCs. Therefore, this study aimed to create the eSCs induced from mouse embryonic stem (mES) cells by regulating defined factors and to explore the relevant regulatory mechanism. Methods Six inducing factors, Sry, Sox9, SF1, WT1, GATA4, and Dmrt1, were respectively transduced into mES cells by lentiviral infection according to the experimental design. The test groups were identified by development stage-specific markers, AMH, Emx2, SF1, and FasL, using flow cytometry. Induced eSCs were determined by FasL and AMH biomarkers under immunofluorescence, immunocytochemistry, and flow cytometry. Moreover, the pluripotency markers, gonad development-related markers, epithelial markers and mesenchymal markers in test groups were transcriptionally determined by qPCR. Results In this study, the co-overexpression of all the six factors effectively produced a large population of eSCs from mES cells in 35 days of culturing. These eSCs were capable of forming tubular-like and ring-like structures with functional performance. The results of flow cytometry indicated that the upregulation of GATA4 and WT1 contributed to the growth of somatic cells in the coelomic epithelium regarded as the main progenitor cells of eSCs. Whereas, SF1 facilitated the development of eSC precursor cells, and Sry and Sox9 promoted the determination of male development. Moreover, the overexpression of Dmrt1 was essential for the maintenance of eSCs and some of their specific surface biomarkers such as FasL. The cellular morphology, biomarker identification, and transcriptomic analysis aided in exploring the regulatory mechanism of deriving eSCs from mES cells. Conclusion Conclusively, we have elucidated a differentiation roadmap of eSCs derived from mES cells with a relevant regulatory mechanism. Through co-overexpression of all these six factors, a large population of eSCs was successfully induced occupying 24% of the whole cell population (1 × 105 cells/cm2). By adopting this approach, a mass of embryonic Sertoli cells can be generated for the purpose of co-culture technique, organ transplantation, gonadal developmental and sex determination researches. |
| first_indexed | 2024-12-12T07:15:34Z |
| format | Article |
| id | doaj.art-fafbf340bb71452ba23d876dc3a5990f |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-12T07:15:34Z |
| publishDate | 2019-03-01 |
| publisher | BMC |
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| series | Stem Cell Research & Therapy |
| spelling | doaj.art-fafbf340bb71452ba23d876dc3a5990f2022-12-22T00:33:31ZengBMCStem Cell Research & Therapy1757-65122019-03-0110111210.1186/s13287-019-1180-6Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cellsChenze Xu0Ali Mohsin1Yanxia Luo2Lili Xie3Yan Peng4Qizheng Wang5Haifeng Hang6Yingping Zhuang7Meijin Guo8State Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyAbstract Background Embryonic Sertoli cells (eSCs) play an important role in sex determination and in male gonad development which makes them a very useful cell type for therapeutic applications. However, the deriving mechanism of Sertoli cells has been unclear and challenging to create a large number of quality eSCs. Therefore, this study aimed to create the eSCs induced from mouse embryonic stem (mES) cells by regulating defined factors and to explore the relevant regulatory mechanism. Methods Six inducing factors, Sry, Sox9, SF1, WT1, GATA4, and Dmrt1, were respectively transduced into mES cells by lentiviral infection according to the experimental design. The test groups were identified by development stage-specific markers, AMH, Emx2, SF1, and FasL, using flow cytometry. Induced eSCs were determined by FasL and AMH biomarkers under immunofluorescence, immunocytochemistry, and flow cytometry. Moreover, the pluripotency markers, gonad development-related markers, epithelial markers and mesenchymal markers in test groups were transcriptionally determined by qPCR. Results In this study, the co-overexpression of all the six factors effectively produced a large population of eSCs from mES cells in 35 days of culturing. These eSCs were capable of forming tubular-like and ring-like structures with functional performance. The results of flow cytometry indicated that the upregulation of GATA4 and WT1 contributed to the growth of somatic cells in the coelomic epithelium regarded as the main progenitor cells of eSCs. Whereas, SF1 facilitated the development of eSC precursor cells, and Sry and Sox9 promoted the determination of male development. Moreover, the overexpression of Dmrt1 was essential for the maintenance of eSCs and some of their specific surface biomarkers such as FasL. The cellular morphology, biomarker identification, and transcriptomic analysis aided in exploring the regulatory mechanism of deriving eSCs from mES cells. Conclusion Conclusively, we have elucidated a differentiation roadmap of eSCs derived from mES cells with a relevant regulatory mechanism. Through co-overexpression of all these six factors, a large population of eSCs was successfully induced occupying 24% of the whole cell population (1 × 105 cells/cm2). By adopting this approach, a mass of embryonic Sertoli cells can be generated for the purpose of co-culture technique, organ transplantation, gonadal developmental and sex determination researches.http://link.springer.com/article/10.1186/s13287-019-1180-6Embryonic stem cellsEmbryonic Sertoli cellsLentiviral transductionMolecular mechanismGonadogenesisMale determinant factors |
| spellingShingle | Chenze Xu Ali Mohsin Yanxia Luo Lili Xie Yan Peng Qizheng Wang Haifeng Hang Yingping Zhuang Meijin Guo Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cells Stem Cell Research & Therapy Embryonic stem cells Embryonic Sertoli cells Lentiviral transduction Molecular mechanism Gonadogenesis Male determinant factors |
| title | Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cells |
| title_full | Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cells |
| title_fullStr | Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cells |
| title_full_unstemmed | Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cells |
| title_short | Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cells |
| title_sort | differentiation roadmap of embryonic sertoli cells derived from mouse embryonic stem cells |
| topic | Embryonic stem cells Embryonic Sertoli cells Lentiviral transduction Molecular mechanism Gonadogenesis Male determinant factors |
| url | http://link.springer.com/article/10.1186/s13287-019-1180-6 |
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