Epigenetic biomarkers to track differentiation of pluripotent stem cells
Summary
Quality control of induced pluripotent stem cells remains a challenge. For validation of the pluripotent state, it is crucial to determine trilineage differentiation potential toward endoderm, mesoderm, and ectoderm. Here, we report GermLayerTracker, a combination of site-specific DNA methylation (DNAm) assays that serve as biomarker for early germ layer specification. CG dinucleotides (CpGs) were identified with characteristic DNAm at pluripotent state and after differentiation into endoderm, mesoderm, and ectoderm. Based on this, a pluripotency score was derived that tracks reprogramming and may indicate differentiation capacity, as well as lineage-specific scores to monitor either directed differentiation or self-organized multilineage differentiation in embryoid bodies. Furthermore, we established pyrosequencing assays for fast and cost-effective analysis. In the future, the GermLayerTracker could be used for quality control of pluripotent cells and to estimate lineage-specific commitment during initial differentiation events. Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.
| Authors | Schmidt M, Zeevaert K, Elsafi Mabrouk MH, Goetzke R, Wagner W |
|---|---|
| Journal | Stem cell reports |
| Publication Date | 2023 Jan 10;18(1):145-158 |
| PubMed | 36460001 |
| PubMed Central | PMC9860076 |
| DOI | 10.1016/j.stemcr.2022.11.001 |