Atcc crl 1730

The cytotoxic effects of Murtilla fruit extract were determined on Human Umbilical Vein Endothelial Cells (HUVEC) ATCC® CRL-1730. Cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM) high glucose (Gibco, Grand Island, NY) supplemented with 10% Bovine Fetal Serum (BFS) and 1% antibiotic (penicillin/streptomycin) (Hyclone, Salt Lake City, UT) at 37°C, 5% CO₂. Cell viability upon treatment with the fruit extract was assessed through the MTT method described by Mossman [18 (link)] with modifications. Briefly, cells were harvested and seeded onto 96-well microtiter plates (Thermo Scientific, Rockford, USA) containing DMEM plus BFS at a density of 5 × 10⁴ cells mL⁻¹ and allowed to adhere for 24 h. After that, the adhered cells were exposed to Murtilla extract at concentrations ranging from 0.001 to 44 μgGAE/mL. After 24 h incubation, viability was determined by adding 5 μg/mL of MTT solution, followed by incubation at 37°C for 2 h. Cells were washed with HBSS 1x (Hanks's Buffer Salt Solution, Hyclone, Salt Lake City, UT), DMSO was added, and the plate was kept at room temperature for 2 h. Final DMSO concentration did not affect cell viability. The succinate dehydrogenase activity was evaluated by color changes and the absorbance was measured at 514 nm using a Biotech Synergy-HT multiscan photometer.

Human umbilical vein endothelial cells (HUVECs) (ATCC CRL-1730) were obtained from the American Type Culture Collection (Rockville, MD, USA) and cultured in low-glucose Dulbecco’s Modified Eagle’s Medium (DMEM) containing 10% fetal bovine serum (FBS, Cegrogen Biotech GmbH, Stadtallendorf, Germany) and 1% penicillin/streptomycin (Biochrom GmbH, Berlin, Germany). Cells were incubated at 37 °C in 5% CO2 in humidified air.
BMSCs were cultured in Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs (ATCC PCS-500-030), containing 10% FBS (Biochrom GmbH), 1% penicillin/streptomycin (Biochrom GmbH), and L-glutamine (4.5 mM) (Biochrom GmbH). Cells were incubated at 37 °C in 5% CO2 in humidified air.

Human umbilical vein endothelial cells (HUVEC, ATCC-CRL-1730™ from ATCC, VA, USA) were cultured in a F–12K medium containing supplementary components following a protocol of ATCC. After the cell confluency reached 80−90 %, HUVEC were seeded into a 96-well plate at the concentration of 50,000 cells/well. The GIL extracts at 0−62.5 μg/mL were treated on HUVEC for 24 h before incubating 14.5 μM peroxynitrite (Merck Ltd., Darmstadt, Germany) for 1 h. The cell viability of HUVEC was measured using the CellTiter 96® AQ_ueous One Solution Cell Proliferation Assay (Promega Corporation, Madison, WI, USA). When cell culture media with viable cells developed a purple color, the color intensity was determined by light absorption at 490 nm.