The role of insulin as a key regulator of seeding, proliferation, and mRNA transcription of human pluripotent stem cells.
Apoptosis
Insulin
Insulin signaling
Pluripotent stem cells
Proliferation
Transcriptional regulation
Journal
Stem cell research & therapy
ISSN: 1757-6512
Titre abrégé: Stem Cell Res Ther
Pays: England
ID NLM: 101527581
Informations de publication
Date de publication:
29 07 2019
29 07 2019
Historique:
received:
24
10
2018
accepted:
30
06
2019
revised:
06
05
2019
entrez:
31
7
2019
pubmed:
31
7
2019
medline:
26
6
2020
Statut:
epublish
Résumé
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. 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. 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. 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.
Sections du résumé
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.
Identifiants
pubmed: 31358052
doi: 10.1186/s13287-019-1319-5
pii: 10.1186/s13287-019-1319-5
pmc: PMC6664730
doi:
Substances chimiques
IGF1R protein, human
0
Insulin
0
RNA, Messenger
0
Receptor, IGF Type 1
EC 2.7.10.1
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
228Subventions
Organisme : Doris Duke Charitable Foundation
ID : 2016097
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK106236
Pays : United States
Organisme : NIH HHS
ID : U01HL107388
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK120565
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK116750
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK116074
Pays : United States
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