title: Loss of tenomodulin results in reduced self-renewal and augmented senescence of tendon stem/progenitor cells.
abstract: Tenomodulin (Tnmd) is a well-known gene marker for the tendon and ligament lineage, but its exact functions in these tissues still remain elusive. In this study, we investigated Tnmd loss of function in mouse tendon stem/progenitor cells (mTSPC) by implicating a previously established Tnmd knockout (KO) mouse model. mTSPC were isolated from control and Tnmd KO tail tendons and their stemness features, such as gene marker profile, multipotential, and self-renewal, were compared. Immunofluorescence and reverse transcriptase-polymerase chain reaction analyses for stem cell-, tenogenic-, osteogenic-, and chondrogenic-related genes confirmed their stemness and lineage specificity and demonstrated no profound differences between the two genotypes. Multipotential was not significantly affected since both cell types differentiated successfully into adipogenic, osteogenic, and chondrogenic lineages. In contrast, self-renewal assays validated that Tnmd KO TSPC exhibit significantly reduced proliferative potential, which was also reflected in lower Cyclin D1 levels. When analyzing possible cellular mechanisms behind the observed decreased self-renewability of Tnmd KO TSPC, we found that cellular senescence plays a major role, starting earlier and cumulating more in Tnmd KO compared with control TSPC. This was accompanied with augmented expression of the cell cycle inhibitor p53. Finally, the proliferative effect of Tnmd in TSPC was confirmed with transient transfection of Tnmd cDNA into Tnmd KO TSPC, which rescued their proliferative deficit. Taken together, we can report that loss of Tnmd affects significantly the self-renewal and senescence properties, but not the multipotential of TSPC.
authors.affiliation: 1 Experimental Surgery and Regenerative Medicine, Department of Surgery, Ludwig-Maximilians-University (LMU) , Munich, Germany .
authors.forename: Paolo
authors.initials: P
authors.lastname: Alberton
authors.forename: Sarah
authors.initials: S
authors.lastname: Dex
authors.forename: Cvetan
authors.initials: C
authors.lastname: Popov
authors.forename: Chisa
authors.initials: C
authors.lastname: Shukunami
authors.forename: Matthias
authors.initials: M
authors.lastname: Schieker
authors.forename: Denitsa
authors.initials: D
authors.lastname: Docheva
mesh_terms: D053687:Adult Stem Cells / Q000502:physiology*; D000818:Animals; D002454:Cell Differentiation; D000066673:Cell Self Renewal; D002478:Cells, Cultured; D016922:Cellular Senescence; D015870:Gene Expression; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D018345:Mice, Knockout; D013710:Tendons / Q000166:cytology*
chemical_list: D008565:Membrane Proteins; C486026:Tnmd protein, mouse