Mesenchymal stem cells (MSCs) raise high hopes in regenerative medicine – but reliable molecular markers to discern MSCs and fibroblasts were so far elusive. Our Epi-MSC-Score provides a simple and robust approach to support the classification of these two cell types. This facilitates quality control of your cell preparations.
How does it work?
To identify relevant genomic regions that can discern MSCs and fibroblasts, we utilized 83 DNA methylation (DNAm) profiles from 12 studies for a training set and 107 DNAm profiles of 16 studies as validation set. Thereby, we identified two CpG dinucleotides – associated with the genes C3orf35 and CIDEC – that are rather methylated in MSCs or fibroblasts, respectively. The Epi-MSC-Score is determined as the difference of these DNAm levels: a level above 0 is indicative for MSCs. We generated pyrosequencing assays that facilitate fast and cost-effective analysis of DNAm levels at the two relevant CpGs (C3orf35 and CIDEC).
Determining the tissue of origin of MSCs
Mesenchymal stem cells (MSCs) can be isolated from various different tissues, such as bone marrow (BM) or adipose tissue (AT). There is a growing perception that MSCs isolated from these two tissues differ significantly in their differentiation potential, hematopoiesis supportive function, and immunomodulatory potential – although they look phenotypically very similar. Our Epi-Tissue-Score enables retrospective analysis of the tissue of isolation. We identified two CpG dinucleotides – associated with the genes SLC41A2 and TM4SF1 – that are either methylated in MSCs from bone marrow or adipose tissue, respectively. The Epi-Tissue-Score is determined as the difference of these DNAm levels: a level above 0 is indicative for MSCs from bone marrow. We generated pyrosequencing assays that facilitate fast and cost-effective analysis of DNAm levels at the two relevant CpGs (SLC41A2 and TM4SF1 ).
Publication:
de Almeida CD et al., Epigenetic Classification of Human Mesenchymal Stromal Cells. Stem Cell Reports 2016.
Patent application: 2016; EP16152198.4