RGD Reference Report - Effect of In-Vitro Passaging on the Stem Cell-related Properties of Tendon-Derived Stem Cells (TDSCs) - Implication in Tissue Engineering. - Rat Genome Database
This study aimed to compare clonogenicity, proliferation, stem cell-related marker expression, senescence, and differentiation potential of rat patellar tendon-derived stem cells (TDSCs) at early (P5), mid (P10) and late (P20, P30) passages. The clonogenicity of the cells was assessed by colony-forming assay while their proliferative potential was assessed by BrdU assay. The surface expression of CD90 and CD73 was assessed by flow cytometry. The cellular senescence was assessed by -galactosidase activity. The adipogenic, chondrogenic and osteogenic differentiation potentials of TDSCs were assessed by standard assays after induction. The mRNA expression of tendon-related markers, Scx and Tnmd, were measured by qRT-PCR. Both the colony numbers and proliferative potential of TDSCs increased with passaging. Concomitantly, there was significant up-regulation of -galactosidase activity with TDSC passaging. The sub-culture of TDSCs downregulated the expression of CD90 and CD73. Lipid droplets were formed in the early and mid passages of TDSCs upon adipogenic induction, but were absent in the late passages. The expression of PPAR2 and C/EBP in TDSCs after adipogenic induction decreased with passaging. Chondrogenesis, proteoglycan deposition, collagen type II protein expression, Col2A1 and Acan mRNA expression were less in pellets formed with later passages of TDSCs after chondrogenic induction. The expression of Scx and Tnmd was lower in the late, compared to early and mid, passages of TDSCs. However, matrix mineralization, Alp and Bglap mRNA expression after osteogenic induction increased with TDSC passaging. Researchers and clinicians should consider the changes of stem cell-related properties of TDSCs when multiplying them in-vitro for tissue engineering.