論文 - 詳細
| RRC ID | 58097 |
|---|---|
| 著者 | Yoda K, Ohnuki Y, Masui S, Kurosawa H. |
| タイトル | Optimized conditions for the supplementation of human-induced pluripotent stem cell cultures with a GSK-3 inhibitor during embryoid body formation with the aim of inducing differentiation into mesodermal and cardiac lineage. |
| ジャーナル | J Biosci Bioeng |
| Abstract |
We optimized the conditions for the differentiation of human induced pluripotent stem cells (hiPSCs) into mesoderm lineage-committed cells by supplementing the cultures with CHIR, a selective GSK-3 inhibitor, during embryoid body (EB) formation. In vitro treatment with 4 μM CHIR during the late 2 days of a 4-day suspension culture period was most effective at promoting mesodermal differentiation. The resulting EBs showed a significant increase in the expression levels of mesoderm-associated genes (WNT3A, T, DKK1, GATA4, FOXC1, and MESP1) and a maintenance of OCT3/4 and NANOG expressions. Upon subsequent differentiation into a cardiac cell lineage, these EBs were shown to generate contractile cardiomyocytes. When shortening the CHIR treatment period to 1 day, the resulting EBs showed reduced expression of mesoderm-associated genes in comparison to the 2-day CHIR treatment. In particular, the expression level of FOXC1 in the 1-day CHIR-treated EBs was much lower than that of the 2-day CHIR-treated EBs. When the treatment period with CHIR was extended to 4 days, the resulting EBs presented significantly reduced expression of WNT3A, OCT3/4, and NANOG upon CHIR concentrations above 4 μM. Similarly, when CHIR treatment was conducted after the formation of EBs, the effectiveness of the GSK-3 inhibitor was reduced compared to a treatment performed during EB formation. Our results indicate that spatiotemporal constraints associated with EB formation, i.e., three-dimensional structuration and cell development in EBs, should be taken into account when designing EB formation-based differentiation protocol involving CHIR treatment. |
| 巻・号 | 129(3) |
| ページ | 371-378 |
| 公開日 | 2020-3-1 |
| DOI | 10.1016/j.jbiosc.2019.09.015 |
| PII | S1389-1723(19)30604-8 |
| PMID | 31615734 |
| MeSH | Animals Cell Differentiation / drug effects* Cell Line Cell Lineage / drug effects Embryoid Bodies / cytology Embryoid Bodies / drug effects* Glycogen Synthase Kinase 3 / antagonists & inhibitors* Humans Induced Pluripotent Stem Cells / cytology Induced Pluripotent Stem Cells / drug effects* Mesoderm / cytology Mesoderm / drug effects* Myocytes, Cardiac / cytology Myocytes, Cardiac / drug effects* Protein Kinase Inhibitors / pharmacology* |
| IF | 2.366 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | 201B7(HPS0063) |