Effects of Long-term Culture on the Biological Characteristics and ceRNA Network-associated RNA Profiles of Human Bone Marrow-derived Mesenchymal Stem Cells

Expansion in vitro prior to mesenchymal stem cells (MSCs) application is a necessary process. Functional and genomic stability has a crucial role in stem cell-based therapies. Long non-coding RNAs (lncRNAs), circular RNAs (circRNAs) and messenger RNAs (mRNAs) play an important role in human bone marrow (BM)-derived MSCs, however, the exact expression and co-expressed profiles in human BM-MSCs in vitro ageing are still lacking. The decreased capacity of proliferation, differentiation and immunoregulation of MSCs at passage (P) 4, P6, P8, P10 and P12 was investigated, and RNA-sequencing identified 439 mRNAs, 65 lncRNAs, 59 miRNAs and 229 circRNAs were differentially expressed (DE) in P12 compared to P4, with a similar trend in P6. GO and KEGG analyses identified several significant biological processes and regulatory pathways, including binding, ossification, and Wnt and PPAR signalling pathways. Interaction and co- expression/co-localization analyses were performed for DE mRNAs and lncRNAs, and several key lncRNAs, circRNAs and important pathway like autophagy and mitophagy were identified in the competing endogenous RNA (ceRNA) network. Our studies indicate replicative senescence of MSC is a continuous process, including widespread alterations in phenotypic, proliferative, differentiation and immunoregulatory potential and global gene expression patterns that need to be considered before therapeutic applications of MSCs. Examination of changes in ceRNA Network-associated RNA profiles of human bone marrow-derived mesenchymal stem cells.