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Determination of the IC50 and LD50 of Calcium Sulfide (CaS) Clusters on Malignant Carcinoma and Normal Fibroblasts Cell Lines (LB580)
Author(s) -
ManoharanValerio Michael,
Ortiz Carlos,
Quiñones Jaileen,
Feliciano Geishamarie,
Diaz Angel,
Rivera Daniel,
Castro Miguel,
Suarez Edu
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.lb580
Subject(s) - trypsinization , cell culture , bioassay , hemocytometer , chemistry , laboratory flask , microbiology and biotechnology , cell counting , andrology , cell , fibroblast , cell growth , calcium , biology , cell cycle , biochemistry , in vitro , medicine , trypsin , genetics , enzyme , organic chemistry
Preliminary results in our lab showed that naked Calcium Sulfide (CaS) nanoparticles (5.0nm) decreased cell proliferation in the carcinoma cell lines CRL‐2124. Objective: The objective of this study was to determine the IC50 and LD50 for CaS clusters (measuring < 3.0 nm) in the carcinoma cell lines CRL‐2124 and the normal fibroblasts CRL‐2522 by using the n3D BiOAssay (Biosciences, Inc.). Methods: A total of 100,000 cells per cell line were seeded in T‐75 flasks. When reaching 80% confluence, they were incubated with the n3D nano‐shuttles overnight (ON) at 37oC, 5% CO2, and 98% relative humidity. Next day, cells were trypsinized, counted, and seeded (1.4 ‐ 1.6 M/per well) in 6‐well plates using the n3D BiOAssay to levitate the cells. A total of 150,000 cells/well were seeded in a 96‐well plate in triplicates with a total volume of 300ul (150ul of cells, 120ul of media, and 30ul of the treatments). The treatments consist of doses ranging 10‐100 pmoles of CaS, 2% DMSO, or growth media. A 24‐hours recording consisting of four (4) photos per minute was collected with an iPod. The analyses were performed using the ImageJ software by measuring the “O” ring outer and inner diameters. Results: We observed changes in the diameters for the wells treated with CaS at different concentrations when compared to controls. Conclusions: We determined the IC50 and LD50 for adenocarcinoma and normal fibroblast cell lines. This step is important for the translation of the use of CaS as an anti‐cancer therapy when translated into in vivo models. Grant Funding Source : NIH‐NIGMSR25GM096955, NIH‐R25GM088023