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Description
Introduction
Stem cell-based therapies are considered one of the most promising disciplines in the field
of biomedicine. Bladder cancer patients could benefit from such therapies directed to promote healing after invasive surgeries or to lessen urinary incontinence, a common side effect of both cancer itself and the treatment. One particularly attractive source of cells for bladder regeneration is adipose tissue. Adipose-derived stem cells (ASCs) of urological patients show similar characteristics, e.g. growth rate, surface markers expression, and differentiation potential to ASCs isolated from healthy individuals. They also secrete a wide variety of soluble mediators that promote tissue regeneration. However, local delivery of ASCs producing large amounts of paracrine factors may promote tumor reactivation.
Methodology
To characterize the interplay between ASCs and bladder cancer cells we used an in-direct co-culture system. ASC52telo mesenchymal stem cells immortalized with hTERT (SCRC-4000) were purchased from ATCC. These primary cells undergo quality control tests to guarantee cell identity. According to International Society for Cell & Gene Therapy (ISCT) criteria ASC52telo cells show high expression of CD29, CD44, CD73, CD90, CD105, CD166 (>90%) and low expression of CD14, CD19, CD34, CD45 (<5%). They retain a multipotent phenotype with the ability to differentiate into adipocytes, osteoblasts. ASCs were seeded on multiwell cell culture plates and bladder cancer cells (5637, HT-1376, and HB-CLS-1) on PET inserts. After 72h co-culture, we analysed morphology, immunophenotype, growth, viability, migration, and activation of intracellular signaling pathways.
Results
Cancer cell-derived factors altered ASCs morphology. Cells with atypical shape and significantly enlarged volume appeared within the monolayer. These morphological changes were not, however, accompanied by changes in the cell surface markers expression. Incubation in conditioned medium (CM) containing soluble mediators secreted by 5637 and HB-CLS-1 cells decreased ASCs number. A significant reduction in viability and proliferation was also noted. In turn, no significant differences were observed when ASCs were co-cultured with HT-1376 cells, which indicates that induced effects may be cell-line dependent.
Conclusions
Adipose-derived stem cells represent one of the most promising cell populations for regenerative medicine. They can be easily harvested in larger quantities and share many of the same regenerative properties as other mesenchymal stem cells. As urological cancer patients are considered good candidates for cell-based therapies, in vitro-expanded ASCs could be used to promote healing after invasive procedures or for organ remodeling surgeries. However, reconstructive cellular therapies for patients with cancer history may promote tumor growth. Our findings indicate that soluble mediators secreted by non-muscle-invasive urinary bladder cancer cells influence ASCs characteristics. For that reason, comprehensive knowledge of the crosstalk between tumor and stem cells seems essential to assess the potential risks of ASC-based regenerative therapies after cancer surgery.
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