Single-cell RNA sequencing reveals metallothionein heterogeneity during hESC differentiation to definitive endoderm
| Autor: | Cheng Zhang, Junjie Lu, Christelle Guillermier, Sean McManus, Hu Li, Anna Baccei, Edroaldo Lummertz da Rocha, Paul H. Lerou, Matthew L. Steinhauser, Lydia Finney |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cell Human Embryonic Stem Cells Intracellular Space Biology 03 medical and health sciences Directed differentiation medicine Metallothionein Humans Induced pluripotent stem cell lcsh:QH301-705.5 Gene Sequence Analysis RNA Endoderm RNA Cell Differentiation Cell Biology General Medicine Cell Cycle Checkpoints Cell biology Zinc 030104 developmental biology medicine.anatomical_structure lcsh:Biology (General) Single-Cell Analysis Pancreas Developmental Biology Definitive endoderm |
| Zdroj: | Stem Cell Research, Vol 28, Iss, Pp 48-55 (2018) |
| ISSN: | 1876-7753 |
| Popis: | Differentiation of human pluripotent stem cells towards definitive endoderm (DE) is the critical first step for generating cells comprising organs such as the gut, liver, pancreas and lung. This in-vitro differentiation process generates a heterogeneous population with a proportion of cells failing to differentiate properly and maintaining expression of pluripotency factors such as Oct4. RNA sequencing of single cells collected at four time points during a 4-day DE differentiation identified high expression of metallothionein genes in the residual Oct4-positive cells that failed to differentiate to DE. Using X-ray fluorescence microscopy and multi-isotope mass spectrometry, we discovered that high intracellular zinc level corresponds with persistent Oct4 expression and failure to differentiate. This study improves our understanding of the cellular heterogeneity during in-vitro directed differentiation and provides a valuable resource to improve DE differentiation efficiency. Keywords: hPSC, Differentiation, Definitive endoderm, Heterogeneity, Single cell, RNA sequencing |
| Databáze: | OpenAIRE |
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