WI‐38 and HFL1 fibroblasts were purchased from Sigma‐Aldrich and cultured in EMEM medium (WI‐38) and HAM's12 medium (HFL1) with 10% fetal bovine serum, 2 mM of L‐glutamine, 1% of non‐essential amino acids and standard Penicillin Streptomycin solution (Sigma‐Aldrich). NHBE epithelial cell line was purchased from Lonza (Lonza Walkersville Inc.) and grown in Bronchial Epithelial Cell Growth Medium BulletKit (Lonza Walkersville Inc.). The experiments (n = 6), were performed after reaching 80%–90% confluence by the cells. The viability of the cells was assessed by adding 10 μL of Presto Blue (BD Pharmingen) and the absorbance was measured at 570 nm.
Presto blue
Manufactured by BD
Sourced in United States
Presto Blue is a cell viability indicator that is used to assess the metabolic activity of cells in cell-based assays. It is a resazurin-based reagent that undergoes a color change when reduced by the metabolic activity of living cells, allowing for the quantification of cell viability and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin solution, by Merck Group (4 mentions)
Wi 38, by Merck Group (4 mentions)
Begm bronchial epithelial cell growth medium bulletkit, by Lonza (3 mentions)
Nhbe epithelial cell line, by Lonza (2 mentions)
Human gingival fibroblasts, by American Type Culture Collection (2 mentions)
Bronchial epithelial cell growth medium bullet kit, by Lonza (1 mentions)
Emem medium, by Merck Group (1 mentions)
6 protocols using presto blue
1
Comparative Fibroblast and Epithelial Cell Culture
2
Comparative In Vitro Evaluation of Respiratory and Gingival Cell Lines
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WI-38 was acquired from (Sigma-Aldrich, St. Louis, MO, USA). The cells were cultured in EMEM medium with 10% fetal bovine serum, 2 mM L-glutamine, 1% non-essential amino acids and standard Penicillin-Streptomycin solution. The NHBE epithelial cell line was obtained from Lonza (Lonza Inc., Walkersville, MD, USA). It was cultivated in BEGM (BulletKit for Bronchial Epithelial Cell Growth Medium) (Lonza Walkersville Inc., Walkersville, MD, USA). Human gingival fibroblasts were purchased from the American Type Culture Collection (ATCC), and cultured in Dulbecco’s Modified Eagle Medium (DMEM), with 10% fetal bovine serum, 100 U/mL penicillin and 0.1 mg/mL streptomycin.
While the use of respiratory cells seems obvious in the context of asthma and airway remodeling, gingival cells were also included to investigate the anti-inflammatory effects of the extracts and to highlight differences between bronchial and gingival epithelia during treatment. All of the experiments (n = 6) were performed after reaching 80–90% confluence (passage three to ten) by the cells. The viability of the cells was evaluated by Presto Blue (BD Pharmingen, Franklin Lakes, NJ, USA), the absorbance was measured at 570 nm.
While the use of respiratory cells seems obvious in the context of asthma and airway remodeling, gingival cells were also included to investigate the anti-inflammatory effects of the extracts and to highlight differences between bronchial and gingival epithelia during treatment. All of the experiments (n = 6) were performed after reaching 80–90% confluence (passage three to ten) by the cells. The viability of the cells was evaluated by Presto Blue (BD Pharmingen, Franklin Lakes, NJ, USA), the absorbance was measured at 570 nm.
3
In Vitro Cell Culture Optimization
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WI-38 were purchased from Sigma-Aldrich (St. Louis, MO, USA) and grown in EMEM medium with 10% fetal bovine serum, 2 mM of L-glutamine, 1% of non-essential amino acids, and standard Penicillin Streptomycin solution (Sigma-Aldrich, St. Louis, MO, USA). The epithelial cell line NHBE was purchased from Lonza (Lonza Walkersville Inc., Walkersville, MD, USA) and cultured in BEGM Bronchial Epithelial Cell Growth Medium BulletKit (Lonza Walkersville Inc., Walkersville, MD, USA). Human Gingival Fibroblasts were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA); the cells were grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/mL, penicillin, and 0.1 mg/mL streptomycin. All the experiments (n = 6) were performed after reaching 80–90% confluence (passage three to ten). The viability of the cells was evaluated by Presto Blue (BD Pharmingen, Franklin Lakes, NJ, USA), and the absorbance was measured at 570 nm.
4
Evaluating Curcumin and Cisplatin Toxicity
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The IC50 values of both curcumin and cisplatin in the A549 and H2170 cells were tested on IMR-90 cells to evaluate the toxic effect of curcumin and cisplatin on normal cells. IMR-90 cells were seeded overnight in 96-well plates at a density of 1.0×104 cells/well in 100 µl complete MEM-α. Subsequently, 100 µl of either curcumin and/or cisplatin (concentration based on IC50 of A549 and H2170) was added to the cells and incubated for 48 h. The viability of the IMR-90 cells was assessed by adding 10 µl of Presto Blue (BD Pharmingen, Franklin Lakes, NJ, USA) to each well and incubated for 2 h before the absorbance was measured at 570 nm.
5
Fibroblast and Epithelial Cell Culture Protocol
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Fibroblast cell lines—WI‐38 and HFL1—were purchased from Sigma‐Aldrich (St. Louis, MO USA) and grown in EMEM medium (WI‐38) and HAM's12 medium (HFL1) with 10% foetal bovine serum, 2 mM of L‐glutamine, 1% of non‐essential amino acids and standard Penicillin‐Streptomycin solution (Sigma‐Aldrich, St. Louis, MO). The epithelial cell line—NHBE—was purchased from Lonza (Lonza Walkersville Inc. MD, USA) and cultured in BEGM Bronchial Epithelial Cell Growth Medium BulletKit (Lonza Walkersville Inc. Walkersville, MD, USA). All the experiments (n = 6) were performed after reaching 80–90% confluence (passage three to nine) by the cells. The viability of the cells was assessed by adding 10 µl of Presto Blue (BD Pharmingen, Franklin Lakes, NJ, USA), and the absorbance was measured at 570 nm.
6
Cell Culture Protocol for Fibroblasts and Epithelium
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WI-38 and HFL1—fibroblast cell lines—were purchased from Sigma-Aldrich (St. Louis, MO, USA). The cells were grown in EMEM medium (WI-38) and HAM’s12 medium (HFL1) with an addition of 10% fetal bovine serum, 2 mM of L-glutamine, 1% of non-essential amino acids, and standard Penicillin Streptomycin solution (Sigma-Aldrich, St. Louis, MO, USA). The epithelial cell line—NHBE—was purchased from Lonza (Lonza Walkersville Inc. Walkersville, MD, USA) and cultured in BEGM Bronchial Epithelial Cell Growth Medium BulletKit (Lonza Walkersville Inc. Walkersville, MD, USA). The experiments (n = 6) were performed after reaching 80–90% confluence (passage three to eight) by the cells. The viability of the cells was assessed using Presto Blue (BD Pharmingen, Franklin Lakes, NJ, USA) and measuring the absorbance at 570 nm.
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