The cervical cancer cell line HeLa, lung cancer cell line NCI-H520 and normal rat kidney epithelial cell line NRK-52E were procured from the National Centre for Cell Science, Pune, India. All the cell lines were grown on 25 cm2 vented culture flasks (Corning, USA). HeLa and NRK-52E cell lines grown using Dulbecco's Modified Eagle Medium (DMEM) (Thermofisher, USA) and NCI-H520 cell line grown using Roswell Park Memorial Institute (RPMI) (Thermofisher, USA) respectively, supplemented with 10% heat-inactivated fetal bovine serum (Thermofisher, USA), and 100 U mL−1 penicillin, 100 μg mL−1 streptomycin and 0.25 μg mL−1 amphotericin B (Thermofisher, USA), in a 37 °C incubator with 5% CO2 and 95% humidity.
25 cm2 vented culture flasks
Manufactured by Corning
Sourced in United States
The 25 cm2 vented culture flasks are a laboratory equipment product designed for cell culture applications. These flasks provide a controlled environment for the growth and maintenance of cells in vitro. The vented design allows for gas exchange between the flask interior and the external environment, supporting the respiratory needs of the cultured cells.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
2 protocols using 25 cm2 vented culture flasks
1
Cultivation of Cell Lines for Cancer Research
2
Optimizing S. rosetta Culture Conditions
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Strains of S. rosetta were co-cultured with Echinicola pacifica bacteria (Levin and King, 2013 (link)); American Type Culture Collection [ATCC], Manassas, VA; Cat. No. PRA-390) in seawater-based media enriched with glycerol, yeast extract, and peptone to promote the growth of E. pacifica that serve as the choanoflagellate prey (Levin and King, 2013 (link); Booth et al., 2018 (link)). We further supplemented this media with cereal grass (King et al., 2009 (link); Fairclough et al., 2010 (link); Carolina Biological Supply Company, Burlington, NC; Cat. No. 132375), which we call high nutrient media (Supplementary file 1 -Table A), as we noticed that this addition promoted S. rosetta growth to a higher cell density (~107 cells/ml [Figure 3—figure supplement 2A ] versus ~106 cells/ml (Booth et al., 2018 (link)). To maintain rapidly proliferating cells in an abundance of nutrients, cultures were diluted 1 in 30 daily or 1 in 60 every two days into 6 ml of high nutrient media in 25 cm2 vented culture flasks (Corning, Oneonta, NY, USA; Cat. No. 430639) and incubated at 22°C and 60% relative humidity. To prevent an overgrowth of bacteria when S. rosetta experienced stress, such as after transfections or during clonal isolation, we cultured S. rosetta in low nutrient media, which is 0.375x high nutrient media (Supplementary file 1 -Table A).
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