title: Replicative senescence of mesenchymal stem cells: a continuous and organized process.
abstract: Mesenchymal stem cells (MSC) comprise a promising tool for cellular therapy. These cells are usually culture expanded prior to their application. However, a precise molecular definition of MSC and the sequel of long-term in vitro culture are yet unknown. In this study, we have addressed the impact of replicative senescence on human MSC preparations. Within 43 to 77 days of cultivation (7 to 12 passages), MSC demonstrated morphological abnormalities, enlargement, attenuated expression of specific surface markers, and ultimately proliferation arrest. Adipogenic differentiation potential decreased whereas the propensity for osteogenic differentiation increased. mRNA expression profiling revealed a consistent pattern of alterations in the global gene expression signature of MSC at different passages. These changes are not restricted to later passages, but are continuously acquired with increasing passages. Genes involved in cell cycle, DNA replication and DNA repair are significantly down-regulated in late passages. Genes from chromosome 4q21 were over-represented among differentially regulated transcripts. Differential expression of 10 genes has been verified in independent donor samples as well as in MSC that were isolated under different culture conditions. Furthermore, miRNA expression profiling revealed an up-regulation of hsa-mir-371, hsa-mir-369-5P, hsa-mir-29c, hsa-mir-499 and hsa-let-7f upon in vitro propagation. Our studies indicate that replicative senescence of MSC preparations is a continuous process starting from the first passage onwards. This process includes far reaching alterations in phenotype, differentiation potential, global gene expression patterns, and miRNA profiles that need to be considered for therapeutic application of MSC preparations.
authors.affiliation: Department of Medicine V, University of Heidelberg, Heidelberg, Germany. wolfgang_wagner@med.uni-heidelberg.de
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wagner
authors.forename: Patrick
authors.initials: P
authors.lastname: Horn
authors.forename: Mirco
authors.initials: M
authors.lastname: Castoldi
authors.forename: Anke
authors.initials: A
authors.lastname: Diehlmann
authors.forename: Simone
authors.initials: S
authors.lastname: Bork
authors.forename: Rainer
authors.initials: R
authors.lastname: Saffrich
authors.forename: Vladimir
authors.initials: V
authors.lastname: Benes
authors.forename: Jonathon
authors.initials: J
authors.lastname: Blake
authors.forename: Stefan
authors.initials: S
authors.lastname: Pfister
authors.forename: Volker
authors.initials: V
authors.lastname: Eckstein
authors.forename: Anthony D
authors.initials: AD
authors.lastname: Ho
mesh_terms: D017667:Adipocytes / Q000737:chemistry; D002454:Cell Differentiation; D002455:Cell Division; D016922:Cellular Senescence; D004260:DNA Repair / Q000235:genetics; D004261:DNA Replication / Q000235:genetics; D020869:Gene Expression Profiling; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics
chemical_list: D035683:MicroRNAs