ctive: The endometrium, the inner lining of the uterine cavity, plays a key role in women’s reproductive health due to its capacity for cyclic regeneration, decidualization, and embryo implantation. Investigating the potential contribution of endometrial stromal cells ( EnSCs) to the pathogenesis of various reproductive disorders requires long-term cultivation of these cells, which is hindered by the replicative senescence of primary cultures. The aim of this work was to establish an immortalized EnSC line. Methods: An EnSC line with constitutive expression of the human telomerase catalytic subunit (hTERT) was generated via lentiviral transduction. The established line was characterized for proliferative activity, cell size, accumulation of lipofuscin granules, senescence-associated activity of β-galactosidase, expression of total histone H3, the cyclin-dependent kinase inhibitor p21, and the secretory profile component PAI-1. Furthermore, the immunophenotype of immortalized EnSCs was assessed using specific antibodies against CD73, CD90, and CD105. The dynamics of stress-induced apoptosis in hTERT-expressing EnSCs were analyzed using Annexin V and DAPI double staining. Results: hTERT expression in EnSCs significantly increased the replicative potential of the cells and delayed the onset of replicative senescence. Importantly, hTERT-expressing EnSCs maintained their immunophenotype and capacity for stress-induced apoptosis throughout the entire culture period. Conclusions: The established immortalized EnSC line represents a valuable tool for fundamental research in reproductive biology.
Melnik et al. (Tue,) studied this question.