Chondrogenic differentiation followed IGFBP3 loss in human endometrial mesenchymal stem cells

Insulin-like growth factor binding protein 3 (IGFBP3) is a multifunctional protein, able either to stimulate the cell growth or to promote apoptosis. In particular, IGFBP3 plays significant role in propagation of stress-induced senescence in human endometrium-derived mesenchymal stem cells (MESCs) (Vassilieva et al., 2020). We undertook CRISPR/Cas9-mediated IGFBP3 knockout in an effort to decelerate stress-induced senescence in MESCs, but, unexpectedly, IGFBP3-knockout MESCs culture acquired chondrocyte-like features, such as cell condensation and aggregation. We revealed that IGFBP3-knockout MESCs completely lost CD73 and CD90 MESCs positive surface markers, and significantly decreased expression of CD105 and CD146 MESCs positive surface markers. In addition, we found IGFBP3-knockout MESCs aggregates positively stained for Alcian Blue. We also detected expression of collagen type II in IGFBP3-knockout MESCs. The obtained results indicate that MESCs lost stemness after IGFBP3-knockout and underwent differentiation toward chondrogenic lineage. Our findings can enlighten IGFBP3 role in regulation of MESCs chondrogenesis. •CRISPR/Cas9 plasmid transfection caused confirmed IGFBP3 gene knockout in MESCs.•In IGFBP3-KO MESCs, the significant changes in surface marker expression were found.•IGFBP3-KO MESCs acquired chondrocyte-like phenotype.•We suppose that IGFBP3 loss in MESCs could facilitate chondrogenic differentiation.•We observed for the first time the chondrogenic differentiation within MESCs culture in response to IGFBP3 knockout.