Early Differentiation Signatures in Human Induced Pluripotent Stem Cells Determined by Non-Targeted Metabolomics Analysis
Abstract
:1. Introduction
2. Experimental Design
2.1. Culturing Human Induced Pluripotent Stem Cells
2.2. Differentiation Induction, Sample Collection, and Cell Counting
2.3. Non-Targeted Metabolomic Using LC-MS Analysis
2.4. Immunofluorescence and Microscopy Imaging
2.5. Statistical Analysis and Data Visualization
3. Results
3.1. The Determination of the Expression of the Pluripotent Marker (OCT3/4)
3.2. Measurement of Extracellular Metabolites and Their Shifts after 48 h
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Medium | Additives | Working Conc. | Code | Pathway | Lineage |
---|---|---|---|---|---|
E6 | DMSO | 0.2 % v/v | T1 | — | Multi-lineages |
E6 | bFGF | 50 ng mL−1 | T2 | FGF signaling | Multi-lineages |
E6 | IWR-1 endo | 2.5 μM | T3 | Wnt signaling | Ectoderm |
E6 | A83-01 | 2.5 μM | T4 | TGF-β signaling | Ectoderm |
E6 | IWP-2 | 10 μM | T5 | Wnt signaling | Ectoderm |
E6 | SB-505124 | 10 μM | T6 | TGF-β signaling | Ectoderm |
E6 | Y-27632 | 10 μM | T7 | ROCK signaling | Multi-lineages |
E6 | CHIR99021 | 10 μM | T8 | GSK-3 signaling | Mesoderm |
E6 | Multi-lineages | ||||
mTeSR Plus | Pluripotency |
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Abdalkader, R.; Chaleckis, R.; Fujita, T. Early Differentiation Signatures in Human Induced Pluripotent Stem Cells Determined by Non-Targeted Metabolomics Analysis. Metabolites 2023, 13, 706. https://doi.org/10.3390/metabo13060706
Abdalkader R, Chaleckis R, Fujita T. Early Differentiation Signatures in Human Induced Pluripotent Stem Cells Determined by Non-Targeted Metabolomics Analysis. Metabolites. 2023; 13(6):706. https://doi.org/10.3390/metabo13060706
Chicago/Turabian StyleAbdalkader, Rodi, Romanas Chaleckis, and Takuya Fujita. 2023. "Early Differentiation Signatures in Human Induced Pluripotent Stem Cells Determined by Non-Targeted Metabolomics Analysis" Metabolites 13, no. 6: 706. https://doi.org/10.3390/metabo13060706