Human mesenchymal stem cell-derived conditioned media (MSC-CM) protects human microvascular endothelial cells from Shiga toxin type 2 cytotoxicity.

VELEZ GUTIERREZ D; FIORE E ; GÓMEZ F; GARCIA M ; IBARRA C; AMARAL M

Mar del Plata

Congreso; Reunión Anual de Sociedades de Biociencia (SAIC SAFE SAB SAP); 2019

Hemolytic uremic syndrome (HUS) is the clinical triad of thrombocytopenia, anemia, and acute renal failure (ARF). HUS is classically associated with Shiga toxin?producing Escherichia coli infection and affects mainly children under 5 years old. Argentina exhibits the highest incidence rate in the world. HUS lacks a specific treatment and many patients develop chronic kidney disease. Recently, mesenchymal stem/stromal cells (MSCs) have been proposed to treat the ARF. MSCs release several antiapoptotic and proliferative mediators that could mitigate the cytotoxic effects caused by Stx2 on renal cells. The objectives of this work were to isolate human MSCs and to analyze if whether the human mesenchymal Stem Cell-Derived Conditioned Media (MSC-CM) would be able to protect human glomerular endothelial cells (HGEC) from the detrimental effects of Stx2. MSCs were isolated by culturing explants of Warthon?s Jelly from human umbilical cord. Then, cells were subcultured and MSC-CM was collected after 24 h of incubation. HGEC were treated with Stx2 (0.5 ng/ml) and in the presence or not of MSC-CM. After 72 h, cell viability was analyzed by neutral red uptake. Cell morphology analysis was evaluated by light microscopy after staining with H&E. Cell counts were performed on four fields and cell area values were obtained using Image J software. Results are expressed as percentage respect to controls (100%). MSC-CM significantly protected in about 30% the HGEC viability (p