Effects of long-term culture on the biological characteristics and 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. However, the exact expression and co-expressed profiles of coding and non-coding RNAs in human bone marrow (BM)-MSCs in vitro aging are still lacking. In the present studies, the change of morphology, immunophenotype, and capacity of proliferation, differentiation, and immunoregulation of MSCs at passage (P) 4, P6, P8, P10, and P12 were investigated. RNA sequencing identified that 439 mRNAs, 65 long noncoding RNAs (lncRNAs), 59 microRNAs (miRNAs), and 229 circular RNAs (circRNAs) were differentially expressed (DE) in P12 compared with P4, with a similar trend in P6. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene set logical processes and pathways, including binding, ossification, tant pathways like autophagy and mitophagy were identified in the competing endogenous RNA (ceRNA) network. Some key RNAs found in the bioinformatics analysis were validated. to be considered before therapeutic applications of MSCs.

Automated summary

Expansion of mesenchymal stem cells (MSCs) in vitro is a necessary process before application, and functional and genomic stability play crucial roles in stem cell therapies. This study investigated the expression and co-expression profiles of coding and non-coding RNAs in human bone marrow MSCs during in vitro aging, revealing significant differences in RNA levels among different passages. Key RNAs identified in bioinformatics analysis were validated, providing important considerations for therapeutic applications of MSCs.