Abstract

Abstract: The sequential culture expansion has been shown to affect the favorable characteristics of human mesenchymal stem/stromal cells (hMSCs), including their immunomodulatory potential. Hereby, we have investigated the immunomodulatory potential of Yazd human testicular Cells (YhTCs) regarding gene expression profile assessment during their lifespan. Morphological examination, immunostaining of vimentin and fibronectin markers, and flow cytometry analysis were used to confirm cultured cells’ MSC properties. Quantitative RT-PCR was used to investigate the target genes, COX-1, TGF-β, IL-6, VCAM-1, and FAS, at different passage numbers (P3, P6, and P9). Our findings confirmed the MSC properties of YhTCs over the lifespan. The results of COX-1 and IL-6 indicated significantly higher expression levels at the late passages (p‑value = 0.03 and p-value = 0.01, respectively). Despite the similar expression pattern, VCAM-1 expression altered significantly at different passages (p-value = 0.0006), but the changes in TGF-β expression levels were non-significant across the lifespan. The YhTCs maintained the main characteristics of MSCs over the sequential culture expansion. Although some of the target genes in the study were sensitive to the culture expansion in the unstimulated YhTCs (IL-6, COX-1, and VCAM1), it seems that FAS and TGF-β need external stimuli or intracellular signals to be induced in YhTCs. © Pleiades Publishing, Ltd. 2024. ISSN 1990-519X, Cell and Tissue Biology, 2024, Vol. 18, No. 4, pp. 390–402. Pleiades Publishing, Ltd., 2024.