Transfected hADSCs cultured in PM or OM for 14 days were assayed for ARS. The culture medium was discarded, and the cells were fixed with 4% paraformaldehyde for 15–20 min and washed with PBS 3 times. ARS solution (ScienCell, San Diego, CA, USA) was prepared in advance and added to the culture plate; the plate was placed in the incubator for 15 min, the staining solution was removed and the sample was washed with PBS solution three times; the PBS was removed, and the samples were placed under a differential microscope to take photos (Leica DMIRB, Germany).
Ars solution
Manufactured by ScienCell
Sourced in United States
ARS solution is a laboratory reagent used for cell viability and proliferation assays. It contains Alizarin Red S, a dye that binds to calcium deposits, allowing for the quantification of mineralized extracellular matrix in cell cultures.
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Lab products found in correlation
6 protocols using ars solution
1
Alizarin Red S Staining of Transfected hADSCs
2
Evaluating Dental Stem Cell Calcification
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To evaluate the calcified nodules formed from human dental stem cells, the ARS staining assay was performed on Days 7 and 14. The hDPSCs were inoculated at a density of 0.5 × 104 cells/well. Over 14 days, hDPSCs were incubated using all experimental disk eluates. The hDPSCs cultured in OM without experimental eluates were used as the control group. The cells were fixed with 4% paraformaldehyde solution for 20 min and stained with 2% ARS solution (ScienCell, Carlsbad, CA, USA) on Days 7 and 14. To stain, 10% cetylpyridinium chromide (Sigma-Aldrich) was applied for 15 min. Each group was measured in six independent experiments.
3
Quantitative Alizarin Red S Assay for Osteogenesis
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40 mM ARS solution from ScienCell was used for this assay. OM-cultured UC-MSCs were fixed by 4% paraformaldehyde for 15 min. To detect calcium deposits from osteo-differentiated cells, samples were treated with 300 μL of ARS solution in a well for 45 min with shaking. Next, the samples were washed five times with distilled water for in order to completely remove the residual dyes. After taking pictures, quantification steps were performed according to the manufacturer’s directions. The absorbance of ARS was read at 405 nm with a plate reader.
4
Evaluating Calcium Nodule Formation in hBMSCs
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The ARS staining assay was used to evaluate the formation of calcium nodules in hBMSCs. Each experimental disk was incubated in osteogenic medium and kept in an incubator at 37 °C and 100% humidity for 7 days, producing a concentration of 5 mg/mL experimental eluate for each tested material. The superficial fluid was purified by 0.20-μm filters (Minisart; Sartorius Stadium Biotech, Göttingen, Germany). hBMSCs were sprayed at a density of 2.0 × 10⁴ cells/well in a 24-well cell culture plate, then incubated for 14 days in the experimental material eluates. Emdogain at an initial concentration of 30 mg/mL was added to groups 2, 4, 6, and 8, and diluted in osteogenic medium to a final concentration of 100 μg/mL. The cells were fixed with a 4% paraformaldehyde and a 2% ARS solution (ScienCell, Carlsbad, CA, USA) for 20 min. The staining was performed for 15 min with 10% cetylpyridinium chromide (Sigma-Aldrich). Finally, an absorbance microplate reader was used to measure the optical density at 560 nm. Each group was measured in sextuplicate.
5
Alizarin Red S Staining of hADSCs
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Transfected hADSCs cultured in PM or OM for 14 days were assayed for ARS. The culture medium was discarded, and the cells were fixed with 4% paraformaldehyde for 15-20 min and washed with PBS 3 times. ARS solution (ScienCell, San Diego, CA, USA) was prepared in advance and added to the culture plate; the plate was placed in the incubator for 15 min, the staining solution was removed, and the sample was washed with PBS solution three times; the PBS was removed and the samples was placed under a differential microscope to take photos (Leica DMIRB, Germany).
6
Alizarin Red Staining of Transfected hADSCs
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Transfected hADSCs cultured in PM or OM for 14 days were assayed for ARS. The culture medium was discarded, and the cells were xed with 4% paraformaldehyde for 15-20 min and washed with PBS 3 times. ARS solution (ScienCell, San Diego, CA, USA) was prepared in advance and added to the culture plate; the plate was placed in the incubator for 15 min, the staining solution was removed, and the sample was washed with PBS solution three times; the PBS was removed and the samples was placed under a differential microscope to take photos (Leica DMIRB, Germany).
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