The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cells
Abstract Background Human-induced pluripotent stem cells (hiPSCs) show a great promise as a renewable source of cells with broad biomedical applications. Since insulin has been used in the maintenance of hiPSCs, in this study we explored the role of insulin in culture of these cells. Methods We repo...
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| Format: | Article |
| Language: | English |
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BMC
2019-07-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13287-019-1319-5 |
| _version_ | 1819054543379365888 |
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| author | Mohammad Shahbazi Paige Cundiff Wenyu Zhou Philip Lee Achchhe Patel Sunita L. D’Souza Fahim Abbasi Thomas Quertermous Joshua W. Knowles |
| author_facet | Mohammad Shahbazi Paige Cundiff Wenyu Zhou Philip Lee Achchhe Patel Sunita L. D’Souza Fahim Abbasi Thomas Quertermous Joshua W. Knowles |
| author_sort | Mohammad Shahbazi |
| collection | DOAJ |
| description | Abstract Background Human-induced pluripotent stem cells (hiPSCs) show a great promise as a renewable source of cells with broad biomedical applications. Since insulin has been used in the maintenance of hiPSCs, in this study we explored the role of insulin in culture of these cells. Methods We report conditions for insulin starvation and stimulation of hiPSCs. Crystal violet staining was used to study the adhesion and proliferation of hiPSCs. Apoptosis and cell cycle assays were performed through flow cytometry. Protein arrays were used to confirm phosphorylation targets, and mRNA sequencing was used to evaluate the effect of transcriptome. Results Insulin improved the seeding and proliferation of hiPSCs. We also observed an altered cell cycle profile and increase in apoptosis in hiPSCs in the absence of insulin. Furthermore, we confirmed phosphorylation of key components of insulin signaling pathway in the presence of insulin and demonstrated the significant effect of insulin on regulation of the mRNA transcriptome of hiPSCs. Conclusion Insulin is a major regulator of seeding, proliferation, phosphorylation and mRNA transcriptome in hiPSCs. Collectively, our work furthers our understanding of human pluripotency and paves the way for future studies that use hiPSCs for modeling genetic ailments affecting insulin signaling pathways. |
| first_indexed | 2024-12-21T12:53:18Z |
| format | Article |
| id | doaj.art-738dfd7f12cf4a188f36d4abaae5f1df |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-21T12:53:18Z |
| publishDate | 2019-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-738dfd7f12cf4a188f36d4abaae5f1df2022-12-21T19:03:25ZengBMCStem Cell Research & Therapy1757-65122019-07-0110111110.1186/s13287-019-1319-5The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cellsMohammad Shahbazi0Paige Cundiff1Wenyu Zhou2Philip Lee3Achchhe Patel4Sunita L. D’Souza5Fahim Abbasi6Thomas Quertermous7Joshua W. Knowles8Stanford Cardiovascular Medicine and Cardiovascular Institute, Stanford School of Medicine, Stanford UniversityDepartment of Developmental and Regenerative Biology, Icahn School of Medicine at Mount SinaiStanford Center for Genomics and Personalized Medicine, Stanford UniversityStanford Cardiovascular Medicine and Cardiovascular Institute, Stanford School of Medicine, Stanford UniversityDepartment of Developmental and Regenerative Biology, Icahn School of Medicine at Mount SinaiDepartment of Developmental and Regenerative Biology, Icahn School of Medicine at Mount SinaiStanford Cardiovascular Medicine and Cardiovascular Institute, Stanford School of Medicine, Stanford UniversityStanford Cardiovascular Medicine and Cardiovascular Institute, Stanford School of Medicine, Stanford UniversityStanford Cardiovascular Medicine and Cardiovascular Institute, Stanford School of Medicine, Stanford UniversityAbstract Background Human-induced pluripotent stem cells (hiPSCs) show a great promise as a renewable source of cells with broad biomedical applications. Since insulin has been used in the maintenance of hiPSCs, in this study we explored the role of insulin in culture of these cells. Methods We report conditions for insulin starvation and stimulation of hiPSCs. Crystal violet staining was used to study the adhesion and proliferation of hiPSCs. Apoptosis and cell cycle assays were performed through flow cytometry. Protein arrays were used to confirm phosphorylation targets, and mRNA sequencing was used to evaluate the effect of transcriptome. Results Insulin improved the seeding and proliferation of hiPSCs. We also observed an altered cell cycle profile and increase in apoptosis in hiPSCs in the absence of insulin. Furthermore, we confirmed phosphorylation of key components of insulin signaling pathway in the presence of insulin and demonstrated the significant effect of insulin on regulation of the mRNA transcriptome of hiPSCs. Conclusion Insulin is a major regulator of seeding, proliferation, phosphorylation and mRNA transcriptome in hiPSCs. Collectively, our work furthers our understanding of human pluripotency and paves the way for future studies that use hiPSCs for modeling genetic ailments affecting insulin signaling pathways.http://link.springer.com/article/10.1186/s13287-019-1319-5InsulinInsulin signalingProliferationApoptosisPluripotent stem cellsTranscriptional regulation |
| spellingShingle | Mohammad Shahbazi Paige Cundiff Wenyu Zhou Philip Lee Achchhe Patel Sunita L. D’Souza Fahim Abbasi Thomas Quertermous Joshua W. Knowles The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cells Stem Cell Research & Therapy Insulin Insulin signaling Proliferation Apoptosis Pluripotent stem cells Transcriptional regulation |
| title | The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cells |
| title_full | The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cells |
| title_fullStr | The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cells |
| title_full_unstemmed | The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cells |
| title_short | The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cells |
| title_sort | role of insulin as a key regulator of seeding proliferation and mrna transcription of human pluripotent stem cells |
| topic | Insulin Insulin signaling Proliferation Apoptosis Pluripotent stem cells Transcriptional regulation |
| url | http://link.springer.com/article/10.1186/s13287-019-1319-5 |
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