Pluripotent - or not pluripotent?
Reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) has revolutionized stem cell research. However, not every cell converts completely into pluripotent state. Our Epi-Pluri-Score provides a simple and robust method for classification of human cells into pluripotent and non-pluripotent cells.
How does it work?
Our epigenetic pluripotency biomarker is based on DNA methylation (DNAm) levels at three specific CpG sites: two CpGs are located within the genes ANKRD46 (methylated in pluripotent cells) and C14orf115 (non-methylated in pluripotent cells) that are combined as Epi-Pluri-Score. The third CpG is located within the pluripotency gene POU5F1 (OCT4) and demarcates early differentiation events. A positive Epi-Pluri-Score indicates pluripotency. Validation of the Epi-Pluri-Score has been performed on 2,215 independent DNA methylation profiles and demonstrated 99,9% specificity and 98,9% sensitivity.
Comparison with conventional pluripotency assays
Several methods can be applied for quality control of human iPSCs, such as teratoma formation, expression of pluripotency-associated genes, or staining of molecular markers (e.g. OCT4, NANOG, TRA-1-60) via immunofluorescence. However, these approaches either require time- and labor-intensive animal experiments or they are not easy to standardize. In contrast, our Epi-Pluri-Score provides a quantitative measure.
Patent application: 2014; EP 14192699.8 (Cygenia GmbH)