Isolated cells (10 × 103) from the deciduous dental pulp tissue were seeded into a 100-mm culture dish (Corning) and were cultured in 10 mL of XFM (Biological Industries) for 14 days. The culture dish was treated with 3 mL of 4% paraformaldehyde (Merck, Darmstadt, Germany) and 0.1% toluidine blue (Merck) in PBS (pH 7.4; Nacalai Tesque) for 18 h. After washing five times with 1 mL of PBS, the dish was air-dried and was imaged with a GT-X980 scanner (Epson, Suwa, Nagano). Numbers of CFU-F, which contained > 50 cells, were counted using a Primovert inverted microscope (Carl Zeiss Microscopy).
100 mm culture dish
Manufactured by Corning
Sourced in United States
The 100-mm culture dish is a laboratory equipment used for cell and tissue culture applications. It provides a sterile, controlled environment for the growth and observation of cells. The dish measures 100 millimeters in diameter and is made of high-quality materials suitable for cell culture processes.
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Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (12 mentions)
Amphotericin b, by Thermo Fisher Scientific (6 mentions)
Tryple, by Thermo Fisher Scientific (6 mentions)
Aec supplement, by PromoCell (6 mentions)
Aec growth medium, by PromoCell (3 mentions)
F12 medium, by Thermo Fisher Scientific (2 mentions)
Amphotericin b, by GE Healthcare (2 mentions)
Penicillin streptomycin, by GE Healthcare (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Complete aec growth medium, by PromoCell (2 mentions)
Toluidine blue, by Merck Group (1 mentions)
Primovert inverted microscope, by Zeiss (1 mentions)
Gt x980 scanner, by Epson (1 mentions)
Crystal violet, by Solarbio (1 mentions)
Hyclone fetal bovine serum, by GE Healthcare (1 mentions)
Human m csf, by R&D Systems (1 mentions)
Il 10, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
16 protocols using 100 mm culture dish
1
Dental Pulp Stem Cell CFU-F Assay
2
Primary NHBE Cell Culture Protocol
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For primary cell culture, we used a previously described protocol (12 (link), 57 (link), 58 (link)). Briefly, NHBE cell monolayer passaging was also performed in a 100-mm culture dish (Corning, Inc.). The culture dish was coated with PureCol (Advanced Biometrics) before seeding cryopreserved NHBE passage 0 (P0) cells, and these cryopreserved cells were thawed in a water bath. The cells were maintained in airway epithelial cell (AEC) growth medium (PromoCell) with AEC supplement (PromoCell), 2% penicillin-streptomycin (Thermo Fisher Scientific), and 1% amphotericin B (Thermo Fisher Scientific) at 37°C in a 5% CO2 incubator. Cells were grown to 90% confluence with a medium change every other day. A confluent monolayer of cells was dissociated with TrypLE (Thermo Fisher Scientific), pelleted, and reseeded into a culture dish containing AEC medium with supplements for passaging. A portion of the cells was stored at −176°C in liquid nitrogen. Cells were passaged up to four times (P4).
3
Colony Formation Assay Protocol
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The third-passage cells were seeded at a density of 200 cells in a 100-mm culture dish (Corning). On day 11, the cells were fixed with 4% paraformaldehyde and stained with 0.5% crystal violet (Solarbio). Colonies containing > 50 cells under microscopy were counted. The percentage of colony-forming efficiency was expressed as the total number of colonies divided by the initial number of cells that were seeded and multiplied by 100 (Huang et al. 2009 (link)).
4
NHBE Cell Monolayer Passaging Protocol
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We used a previously described protocol (12 (link), 58 (link), 116 (link)). Briefly, NHBE cell monolayer passaging was also performed in a 100 mm culture dish (Corning, Inc.). The culture dish was coated with PureCol (Advanced Biometrics) before seeding cryopreserved NHBE passage zero (P0) cells, and these cryopreserved cells were thawed in a water bath. The cells were maintained in airway epithelial cell (AEC) growth medium (PromoCell) with AEC supplement (PromoCell), 2% penicillin/streptomycin (Thermo Fisher Scientific) and 1% amphotericin B (Thermo Fisher Scientific) at 37 °C in a 5% CO2 incubator. Cells were grown to 90% confluency with a medium change every other day. A confluent monolayer of cells was dissociated with TrypLE (Thermo Fisher Scientific), pelleted, and reseeded into a culture dish containing AEC medium with supplements for passaging. A portion of the cells was stored at −176°C in liquid nitrogen. Cells were passaged up to four times (P4).
5
Characterization of Au-NPs Uptake in A549 and 95D Cells
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The uptake of Au-NPs was also examined using TEM. Before exposure to the Au-NPs, the A549 cells and 95D cells were plated at a concentration of 1×106 cells per dish on a 100-mm culture dish (corning, USA) containing the growth medium and incubated at 37°C with 5% CO2 for 24 h. The Au-NPs were then added. After exposure to Au-NPs for 48 h, the cells were fixed in 3.7% (v/v) paraformaldehyde in phosphate-buffered saline (PBS) for 20 min at RT. Subsequently, cells were prepared for TEM analysis as follows: cells were fixed in 1% (w/v) osmium tetroxide for 2 h, dehydrated in a graded series of 30%, 50%, 70%, 80%, and 90% ethanol, and treated three times with 100% ethanol for 15 min each. The samples were then embedded in a mixture of resin in propylene oxide polymerized at 80°C. Ultrathin sections for TEM were prepared using a diamond knife and the samples were analyzed using a transmission electron microscope.
6
Differentiation of Primary NHBE Cells
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We passaged the primary NHBE cells (passage 1) three times before differentiating them (passage 4) into a pseudostratified epithelium. For each passage, the cells were grown in a 100-mm culture dish (Corning Inc.) precoated with PurCol (Advanced Biometrics). The cells were maintained in airway epithelial cell (AEC) growth medium (PromoCell) containing AEC supplements (PromoCell), 2% penicillin/streptomycin (Thermos Fisher Scientific), and 1% amphotericin B (Thermos Fisher Scientific) (complete AEC medium) at 37°C in an incubator with 5% CO2. The cells were grown to 90% confluency, and the medium was changed every other day. Confluent monolayers of cells were dissociated with 5 ml of TrypLE (Thermo Fisher Scientific) and pelleted, and one-third of the cells were reseeded in a culture dish containing complete AEC medium for passaging. A549 cells were grown in F-12 medium (Thermo Fisher Scientific) with 10% HyClone fetal bovine serum (GE Healthcare), 2% penicillin/streptomycin, and 1% amphotericin B.
7
Differentiation of M2 Macrophages
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Generation of M2 macrophages was performed according to the protocol described by Leidi et al. [15 (link)]. Briefly, human PBMCs were co-cultured in 6-well plates or 100-mm culture dish (Corning) in complete RPMI1640 media supplemented with 30 ng/ml human M-CSF (R&D systems) for 4 days. Adherent cells were retained by gently washing off non- and loose-adherent cells, with half of media replaced, and culture for 2–3 more days. For M2 polarization, 10 ng/ml IL-10 (Peprotech) was added during the last 48 h of culture.
8
Culturing Human Cancer Cell Lines
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The human cancer cells QBC9939, MZ-Cha-1 were obtained from the American Type of Culture Collection (ATCC, Rockville, MD, USA). They were separately cultured in 100-mm culture dish (Corning Glass Works, Corning, NY, USA) using RPMI-1640 and DMEM medium (Gibco, Grand Island, NY, USA) with 5% fetal bovine serum (Gibco), 100 μg/ml streptomycin and 100 U/ml penicillin at 37°C in 5% CO2 humidified incubator.
9
Isolation and Culture of Human BMSCs
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Human mandibular bones of BRONJ patients or healthy donors were collected at the Peking University Hospital of Stomatology, approved by the Ethics Committee of Peking University (IRB00001052-11002). After surgery, bone biopsies were thoroughly cut into 1-mm3 cubes and digested with dispase II (4 mg/mL) and collagenase I (2 mg/mL) for 30 min at 37°C. Cells were collected by centrifugation at 1,200 rpm for 5 min, then filtered, resuspended and seeded in a 100-mm culture dish (Corning, NY, United States). Following overnight culture at 37°C in a humidified atmosphere of 5% CO2, unattached cells were discarded and the medium was changed every 2 days. Cells were maintained at 37°C and 5% CO2. BMSCs at passage 3 were used for the migration assay.
10
Isolation and Culture of Human Rotator Cuff Tenocytes
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Human rotator cuff tendon was obtained during elective surgery of rotator cuff repair from a 63-year-old woman. Informed written consent was taken from the patient for this study.
The sample, 5 × 5 mm out of a 30 × 20 mm-full thickness of rotator cuff tear, was washed with phosphate buffered saline (PBS: HyClone, USA) three times, and then cut into 1-2 mm 3 with a No. 11 knife. Pieces of torn rotator cuff tissue were placed on cover slides and incubated at 37°C in a CO 2 incubator for two weeks. Cell suspension was collected, filtered through a 100-μm cell strainer (BD Biosciences, USA), and centrifuged at 300 g for 10 min at room temperature. Suspension was incubated with 1 ml of red blood cell lysing buffer (Sigma R7757, USA) at room temperature for 1 min, pipetted for 1 min to mix, added to 9 ml of PBS, and then centrifuged at 300 ×g for 10 min at room temperature. Isolated cells were seeded into a 100-mm culture dish (Corning, USA) and incubated in Dulbecco ' s modified Eagle medium (DMEM: HyClone, USA) containing 10% fetal bovine serum (Gibco, USA), 100 U/ml penicillin and 100 U/ml streptomycin (Gibco). The cultures were maintained at 37°C CO 2 incubator. Culture medium was replaced three times per week. All experiments on tenocytes were conducted at three passages.
The sample, 5 × 5 mm out of a 30 × 20 mm-full thickness of rotator cuff tear, was washed with phosphate buffered saline (PBS: HyClone, USA) three times, and then cut into 1-2 mm 3 with a No. 11 knife. Pieces of torn rotator cuff tissue were placed on cover slides and incubated at 37°C in a CO 2 incubator for two weeks. Cell suspension was collected, filtered through a 100-μm cell strainer (BD Biosciences, USA), and centrifuged at 300 g for 10 min at room temperature. Suspension was incubated with 1 ml of red blood cell lysing buffer (Sigma R7757, USA) at room temperature for 1 min, pipetted for 1 min to mix, added to 9 ml of PBS, and then centrifuged at 300 ×g for 10 min at room temperature. Isolated cells were seeded into a 100-mm culture dish (Corning, USA) and incubated in Dulbecco ' s modified Eagle medium (DMEM: HyClone, USA) containing 10% fetal bovine serum (Gibco, USA), 100 U/ml penicillin and 100 U/ml streptomycin (Gibco). The cultures were maintained at 37°C CO 2 incubator. Culture medium was replaced three times per week. All experiments on tenocytes were conducted at three passages.
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