Cytotoxic effect of MNPs was additionally determined using TPB assay after culturing HeLa cells on 35 mm culture dishes (Corning, USA). This assay was also used to determine cell viability subsequent to all hyperthermic effects on HeLa cells. Briefly, the cells were seeded at a density of 2.5 * 105 cells/culture dish and treated for the time and doses as mentioned in MTT assay above. However, at the end of treatment, instead of adding MTT, the cells, after PBS wash and trypsinization by 0.25% Trypsin-EDTA (Thermofisher, USA), were stained with trypan blue solution and placed on hemocytometer for viability count under the microscope.
35 mm culture dishes
Manufactured by Corning
Sourced in United States
The 35 mm culture dishes are a type of laboratory equipment used for the cultivation and study of cells, tissues, or microorganisms. These petri-style dishes provide a controlled environment for cell growth and experimentation. The dishes are made of transparent, sterile material, typically polystyrene, and have a diameter of 35 millimeters.
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Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Foetal bovine serum fbs, by Greiner (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Esgro, by Merck Group (1 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Poly d lysine, by Merck Group (1 mentions)
Co2 water jacketed incubator, by NuAire (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin, by Merck Group (1 mentions)
Collagenase, by Merck Group (1 mentions)
Hepes, by Merck Group (1 mentions)
Fungizone, by Thermo Fisher Scientific (1 mentions)
Nahco3, by Merck Group (1 mentions)
Leibovitz l 15 medium, by Thermo Fisher Scientific (1 mentions)
α mem, by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
21 protocols using 35 mm culture dishes
1
Trypan Blue Assay for MNP Cytotoxicity
2
Murine Embryonic Stem Cell Maintenance
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Murine embryonic stem cells (ES-D3 (D3), ATCC® (Manassas, VA, USA)) were maintained according to the previously described protocol (Mennen et al., 2019 , Spielmann et al., 1997 ). The embryonic stem cells (ESCs) were maintained in 35 mm culture dishes (Corning, New York, NY, USA) in a humidified atmosphere at 37 °C with 5 % CO2 for stimulation of cell proliferation.. ESCs were replated in fresh medium every 2–3 days. The culture medium (CM) consisted of Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Waltham, MA, USA), 20 % Foetal Bovine Serum (FBS; Greiner Bio-One, Kremsmünster, Austria); 2 mM l -Glutamine (Gibco); 1 % Non-Essential Amino Acids (NEAA; Gibco); 1 % 5000 IU/ml Penicillin/5000 µg/ml Streptomycin (Gibco); and 0.1 mM β-mercaptoethanol (Gibco). In order to preserve pluripotency, the ESCs in CM were supplemented with 1000 units/ml leukemia inhibitory factor (LIF; ESGRO®, Millipore, Burlington, MA, USA). These pluripotent ESCs were used in the differentiation assays.
3
Isolation and Culture of Rat Dorsal Root Ganglia Neurons
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The DRG neurons were isolated from the vertebral column of Wistar rats at postnatal ages P5 to P9 without sex distinction, and cultured according to the procedure described by Salceda and coworkers [71 (link)]. In brief, the neurons were incubated (30 min at 37°C) in Leibovitz L15 medium (Invitrogen, USA) containing 1.25 mg/ml trypsin and 1.25 mg/ml collagenase (both from Sigma-Aldrich). After the enzymatic treatment, the ganglia were washed 3 times with sterile L15. Cells were mechanically dissociated and plated on 12-mm x 10-mm glass coverslips (Corning, USA), pretreated with poly-D-lysine (Sigma-Aldrich), which were placed onto 35-mm culture dishes (Corning). The isolated cells were allowed to settle and adhere to the coverslips during 4 to 8 h in a humid atmosphere (95% air and 5% CO2, at 37°C) using a CO2 water-jacketed incubator (Nuaire, USA). The plating medium was L15 supplemented with 15.7 mM NaHCO3 (Merck, Mexico), 10% fetal bovine serum, 2.5 μg/ml fungizone (both from Invitrogen), 100 U/mL penicillin (Lakeside, Mexico), and 15.8 mM HEPES (Sigma-Aldrich).
4
Wound Healing Assay for GBM10 Cells
5
Population Doubling Time of Cell Cultures
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The population doubling time determines the dynamics of the cell culture development as the average time required for a cell to complete the cell cycle. In the case of cancer cells, population doubling time allows evaluation of the compounds’ efficiency. In the case of increased growth of the cell culture, the compound has a regenerative potential called cell self-renewal.
A total of 3 × 105 A375, C32 and HaCaT cells were seeded in 35 mm culture dishes (Corning, New York, NY, USA). The cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2. After 24 h, the culture supernatants were removed, and appropriate dilutions of compounds in the culture medium were added and incubated for an additional 24 h or 72 h. The cells were collected using trypsin and counted using KOVA (KOVA® Glasstic Slide 10 with Grid Chamber, HYCOR Biomedical, Garden Grove, CA, USA) after 24 and 72 h.
A total of 3 × 105 A375, C32 and HaCaT cells were seeded in 35 mm culture dishes (Corning, New York, NY, USA). The cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2. After 24 h, the culture supernatants were removed, and appropriate dilutions of compounds in the culture medium were added and incubated for an additional 24 h or 72 h. The cells were collected using trypsin and counted using KOVA (KOVA® Glasstic Slide 10 with Grid Chamber, HYCOR Biomedical, Garden Grove, CA, USA) after 24 and 72 h.
6
Murine Bone Marrow Colony Assay
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Murine non-adherent bone marrow cells (1 × 105cells/culture) were cultured in α-MEM (Gibco, Grand Island, NY) containing 1.2% methylcellulose, 30% FBS, 1% deionized bovine serum albumin (BSA) (Sigma-Aldrich), and 100 ng/mL recombinant human GM-CSF (Immunex Corp., Seattle, WA, USA). The cells were plated in a volume of 1.0 mL in 35-mm culture dishes (Corning, New York, NY), as reported previously [31 (link)], and treated with or without Aplidin (5 pM and 50 pM). The number of colonies that formed were scored after 7 days of culture using an inverted microscope.
7
PGC Transplantation in Drosophila Embryos
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PGC transplantation was conducted at 25 °C as previously described18 (link),29 (link). Freeze-thawed PGCs were transplanted into the posterior pole of either y w host embryos or OvoA_OE host embryos at the cellular blastoderm stage (100–150 min AEL). Between 10 and 20 PGCs were injected into each host embryo. The number of PGCs transplanted into host embryos was counted under a microscope. Transplanted embryos were maintained at 25 °C until hatching. Hatched larvae were transferred into a standard culture medium in 35 mm culture dishes (Falcon brand, Corning Life Sciences, Schiphol-Rijk, Netherlands), and incubated at 25 °C until pupation (<15 larvae per dish). The resultant pupae were transferred into vials containing culture medium and maintained at 25 °C until eclosion.
8
Culturing Human Nasal Epithelial Cells
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HNPCECs were purchased from Science Cell Research Laboratories (Carlsbad, CA, USA) and cultured in Dulbecco’s modified Eagle medium (DMEM; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% newborn calf serum (NCS; Invitrogen), 100 U/mL penicillin, and 100 μg/mL streptomycin (Roche Diagnostics, Mannheim , Germany) at 37°C in a humidified atmosphere containing 5% CO2. After the cells reached confluence, cells were harvested with 0.025% trypsin (Invitrogen) in PBS and transferred to plastic culture dishes at a 1:4 split ratio. For experiments, the cells were trypsinized and seeded at 1 × 106 cells/mL in 35-mm culture dishes (Corning Inc., Corning, NY, USA). HNPCECs reached confluence after 72 hr (set as day 0). The HNPCECs from three different donors were used after the third to sixth passages in this study. Results among the three donors were indistinguishable.
9
Embryonic UV-Induced DNA Damage
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To induce DNA damage, embryos at 36 h, D3, D5 or D7 of development post-PA were placed in 35 mm culture dishes (Corning, Tewksbury, MA, United States) containing 1 ml of PZM-3 medium placed in biologic safety cabinet (1300 Series Class II; Thermo Fisher Scientific, Waltham, MA, United States), and then exposed (UV+) or not (UV-) to UV light for 10 s. After UV treatment, embryos were washed and cultured for 30 min, 6 h or up to D7 and collected to extract mRNA for RT-qPCR analyses, assess DNA damage, or evaluate development to the blastocyst stage, depending on the experiment.
10
Immortalized Human Bronchial Epithelial Cell Culture
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Human bronchial epithelial (hBE) cells were immortalized following transfection of genes encoding cyclin-dependent kinase-4 and human telomerase reverse transcriptase [67 (link)]. Cell monolayers were grown on two-well chamber slides (Laboratory-Tek, VWR International, Chicago, IL, USA) for Ca2+ imaging and dye uptake experiments, or on 35 mm culture dishes (Corning Life Sciences, Lowell, MA, USA) for ATP release measurements and for comet assays. The cells were cultured in bronchial epithelial cell growth medium with growth factor supplements (PromoCell GmbH, Heidelberg, Germany) and incubated at 37 °C in a humidified atmosphere of 5% CO2 in air.
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