DOI: 10.6084/m9.figshare.30147294.v1
title: hPL-conditioned MSC-derived extracellular vesicles exhibit enhanced anticancer effects on MCF7 and A549 cells

description: Mesenchymal stem cells (MSCs) have therapeutic potential due to their differentiation and immune-regulatory capabilities, mainly through the secretion of extracellular vesicles (EVs) which can exhibit pro- and anti-tumor effects. This study aimed to investigate the effects of EVs derived from Whorton’s jelly derived MSCs (WJMSCs) cultured in fetal bovine serum (FBS) or human platelet lysate (hPL), as xeno-free alternative, on breast (MCF7) and lung (A549) cancer cells. EVs were isolated from WJMSCs cultured in either FBS or hPL and then applied to cancer cells. Following characterization of EVs, the effects on cancer cell proliferation, apoptosis, senescence, and migration were also assessed. Both EV types reduced proliferation in MCF7 and A549 cells at 24 hours, with a sustained effect on MCF7 at 48 hours. Senescence marked by upregulation of the Senescence-associated β-galactosidase (SA-β-gal) was induced in both cell lines, indicating non-apoptotic growth inhibition. WJMSC-hPL-EVs enhanced migration in MCF7 cells despite reduced proliferation, whereas A549 cell migration was time-dependent, as demonstrated by the wound scratch assay. While the findings show minimal differences between FBS and hPL, the results nonetheless support the potential use of hPL as a viable xeno-free alternative to FBS. Further research is needed to the therapeutic implications. EVs from FBS- and hPL-cultured WJMSCs exhibited similar characteristics, including marker expression, morphology, and size.Both EV types reduced proliferation in MCF7 and A549 cells at 24 hours, with sustained effects in MCF7 cells at 48 hours possibly linked to SA-β-gal upregulation.WJMSC-hPL EVs enhanced migration in MCF7 cells despite reduced proliferative capacity.WJMSC-hPL EVs induced early-stage migration in A549 cells, while a significant inhibition was observed at later time points, indicating a time-dependent effect. EVs from FBS- and hPL-cultured WJMSCs exhibited similar characteristics, including marker expression, morphology, and size. Both EV types reduced proliferation in MCF7 and A549 cells at 24 hours, with sustained effects in MCF7 cells at 48 hours possibly linked to SA-β-gal upregulation. WJMSC-hPL EVs enhanced migration in MCF7 cells despite reduced proliferative capacity. WJMSC-hPL EVs induced early-stage migration in A549 cells, while a significant inhibition was observed at later time points, indicating a time-dependent effect. Breast and lung cancers remain leading causes of death worldwide, highlighting the urgent need for innovative therapies. Mesenchymal stem cells (MSCs), including those derived from the umbilical cord, release small vesicles called extracellular vesicles (EVs) that mediate intracellular communication between cells, including interaction with cancer cells. In this study, we cultured MSCs using two different supplements: fetal bovine serum (FBS), sourced from cow blood and traditionally used in cell culture, and human platelet lysate (hPL), a human-derived alternative with fewer ethical concerns. We then examined how EVs from MSCs grown in these two different conditions influenced breast (MCF7) and lung (A549) cancer cells in the laboratory. Our results showed that EVs from both culture conditions reduced cancer cell proliferation and induced cellular growth arrest without increasing cell death. Notably, EVs from MSCs cultured with hPL enhanced the motility of breast cancer cells, a factor that may affect cancer spread. Lung cancer cells displayed variable migration patterns over time. Overall, the EVs’ effects were broadly similar regardless of the culture supplement used. These findings support the use of hPL as an ethical and effective alternative to animal-derived serum for producing MSC EVs. Nevertheless, further research is needed to fully understand the mechanisms underlying these effects and to evaluate their safety and therapeutic utility in cancer treatment.