T 75 flasks

Manufactured by BD

Sourced in United States

The T-75 flasks are a type of tissue culture flask commonly used in cell culture applications. These flasks have a working volume of 75 cm^3 and provide a surface area for cell growth and manipulation. The flasks are made of transparent, sterile polystyrene material and are equipped with a vented cap to facilitate gas exchange.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Trizol ls reagent, by Thermo Fisher Scientific (1 mentions) Horse serum, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by American Type Culture Collection (1 mentions) F 12k medium, by American Type Culture Collection (1 mentions) Tc10 automated cell counter, by Bio-Rad (1 mentions) Du145 human prostate epithelial cells, by American Type Culture Collection (1 mentions) A549 human lung epithelial cells, by American Type Culture Collection (1 mentions) Opticell, by Thermo Fisher Scientific (1 mentions) Egm 2, by Lonza (1 mentions) Huvecs, by Lonza (1 mentions) Trypsin, by Lonza (1 mentions) Ethylene diamine tetraacetic acid (edta), by Lonza (1 mentions)

Close spelling variants of this product

T-75 cell culture flask T-75 tissue culture flasks 75-cm2 T-flasks T-flasks

7 protocols using t 75 flasks

RNA Extraction from Infected Caco-2 Cells

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Samples were collected as previously described32 (link)34 (link)58 (link)59 (link). Briefly, Caco-2 cells were cultured in T-75 flasks (BD) and were serum starved 24 h before infection. Respective bacterial treatments with Salmonella WT and the deletion mutants, at MOI of 1000, were used to infect epithelial cells as described previously. All treatments were incubated at 37 °C with 5% CO₂ for 60 min. 10 ml of TRIzol LS reagent (Invitrogen, Carlsbad, CA) was added to the cells and mixed with pipette followed by centrifugation at 7,200 × g for 5 min to pellet the host associated bacteria. TRIzol LS supernatant was stored in a clean tube and further processed for RNA extraction from infected Caco-2 cells. The bacterial pellet was suspended in 2 ml of fresh TRIzol LS, gently mixed and further processed for RNA extraction from host associated bacteria. The experiment was done in two biological replicates.

Arabyan N., Park D., Foutouhi S., Weis A.M., Huang B.C., Williams C.C., Desai P., Shah J., Jeannotte R., Kong N., Lebrilla C.B, & Weimer B.C. (2016). Salmonella Degrades the Host Glycocalyx Leading to Altered Infection and Glycan Remodeling. Scientific Reports, 6, 29525.

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Culture and Maintenance of Rodent Adrenal Cells

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Rodent Y1 adrenal cortical cells (ATCC, Manassas, VA) were grown in F12K medium (ATCC) supplemented with 2.5% fetal bovine serum (ATCC), 15% horse serum (Fisher Scientific, St-Louis, MO), and 1% penicillin/streptomycin (Thermo Scientific, Rockford, IL). They were grown as a monolayer in a humidified atmosphere at 37°C in 5% CO₂ in T75 flasks (BD Biosciences, San Jose, CA). The medium was changed every 4 days, and cells were sub-cultured after 8 days and split into a 1:3 ratio. They were then stored in liquid nitrogen (5% dimethyl sulfoxide (DMSO) growth medium) or plated for experiments. All experiments were conducted in passage 4 - passage 6 cells. Cells were counted using the TC10™ Automated Cell Counter (Bio-Rad Life Science, Hercules, CA) and seeded in 12-well plates at a concentration of 1 X 10⁶ live cells/well. The cells were grown in 2ml of growth media/well for 48 hours. They were then serum-starved for 8 hours, as previously described (Calejman et al, 2011 (link)), and subsequently incubated for 24 hours with growth medium containing the appropriate chemicals for the respective groups.

Charles M.S., Perera P.N., Doycheva D.M, & Tang J. (2015). Granulocyte-Colony Stimulating Factor activates JAK2/PI3K/PDE3B Pathway to Inhibit Corticosterone Synthesis in a Neonatal Hypoxic-ischemic Brain Injury Rat Model. Experimental neurology, 272, 152-159.

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Isolation and Culture of Rat Bone Marrow Stromal Cells

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BMSCs were collected from the bone marrow of the rats. Briefly, both ends of the femurs were cut off by sterile operation scissors, and the bone marrow was rinsed thoroughly by PBS containing 1% penicillin-streptomycin (Invitrogen, Carlsbad, CA, USA). After that, the mixture was centrifuged at 500× g for 20 min, and the cell pellet was diluted with Dulbecco’s Modified Eagle’s Media (Hyclone, Logan, UT, USA) containing 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) and 1% penicillin-streptomycin. The cells were cultured in T-75 flasks (BD Biosciences, San Jose, CA, USA) at 37°C in a humidified incubator containing 5% CO₂, and the culture media were changed twice a week. The BMSCs were passaged when they were confluent, and the cells obtained at the end of the third passage were used for the experiments.

Li Y, & Zhang Z.Z. (2018). Sustained curcumin release from PLGA microspheres improves bone formation under diabetic conditions by inhibiting the reactive oxygen species production. Drug Design, Development and Therapy, 12, 1453-1466.

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Isolation and Expansion of Lymphokine-Activated Killer Cells

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Spleens were harvested into single cell suspensions by pulverization through a 70 micron filter as previously described (35 (link)). After lysis of red blood cells, cell suspensions were washed in pre-warmed (RT) HBSS/10% FBS and loaded onto a nylon wool column prepared in a 10 ml syringe. Cells were incubated on the column at 37°C for 1 hr. Non-adherent cells were passed through the column after the addition HBSS/10% FBS, collected and grown in T-75 flasks (BD Biosciences) in the presence of IL-2 (8000 U/mL; Proleukin, Chiron). After 4 days, non-adherent cells were removed and flasks were washed with PBS to remove loosely adherent cells. Fresh IL-2 containing media was replenished in the original flasks. Washes were collected with the non-adherent cells, centrifuged and transferred to a second flask with IL-2 containing media. Cells were maintained for an additional 3–4 days before use. This protocol yielded LAK cells in which 60–90% of the cells were NK1.1⁺, CD3⁻.

Miner C.A., Giri T.K., Meyer C.E., Shabsovich M, & Tripathy S.K. (2015). Acquisition of activation receptor ligand by trogocytosis renders natural killercells hyporesponsive. Journal of immunology (Baltimore, Md. : 1950), 194(4), 1945-1953.

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Cell culture protocol for prostate, lung, and colonic epithelial cells

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DU145 human prostate epithelial cells and A549 human lung epithelial cells were acquired from American Type Culture Collection (Manassas, VA). SK-CO15 human colonic epithelial cells were provided by Dr. Enrique Rodriguez-Boulan (Cornell University). DU145 cells were cultured in RPMI medium supplemented with 10% Fetal Bovine Serum, 15% HEPES, pyruvate, and antibiotics. A549 and SK-CO15 cells were cultured in DMEM/F12 and DMEM medium, respectively, supplemented with 10% Fetal Bovine Serum and antibiotics. The cells were grown in T75 flasks (BD Biosciences), and were seeded on collagen-coated coverslips or 6-well plastic plates for immunolabeling and biochemical experiments, respectively.

Wang D., Chadha G.K., Feygin A, & Ivanov A.I. (2015). F-actin binding protein, anillin, regulates integrity of intercellular junctions in human epithelial cells. Cellular and molecular life sciences : CMLS, 72(16), 3185-3200.

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Isolation and Characterization of Human ADSCs

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Human-ADSCs were purchased from the Type Culture Collection of the Chinese Academy of Sciences. The eight donors (men, 4; women, 4) were aged from 21–42 years old (the sex and age range of each sample were provided by the Type Culture Collection of the Chinese Academy of Sciences). The cells were centrifuged at 500 × g for 20 min at room temperature, and the cell pellet was diluted with D10 media which was composed of 89% DMEM-low glucose (DMEM-LG; Hyclone; Cytiva), 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.). Then, cells were cultured in T-75 flasks (BD Biosciences) at 37°C in a humidified incubator containing 5% CO₂, and the culture media was changed twice a week. ADSCs were passaged when they were confluent, and the cells obtained at the end of the third passage were used for the experiments. ADSCs from different donors were separately cultured. ADSCs from all eight donors were used in all subsequent experiments, including the animal study.

Li R., Li Y, & Dong X. (2020). Preconditioning mesenchymal stromal cells with flagellin enhances the anti-inflammatory ability of their secretome against lipopolysaccharide-induced acute lung injury. Molecular Medicine Reports, 22(4), 2753-2766.

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Culturing Human Umbilical Vein Endothelial Cells

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Human umbilical vein endothelial cells (HUVECs) (Lonza, Verviers, Belgium) were cultured in EGM-2 (Lonza) medium in T75 flasks (BD, Breda, the Netherlands) in a humidified incubator at 37°C with 5% CO₂. Cells were detached with 0.25% Trypsin in EDTA (Lonza) and replated on one side of the acoustically transparent OptiCell^™ (NUNC, Wiesbaden, Germany) chambers. HUVECs were cultured as described before [48 (link)], for two days until 70% confluence to resemble neovasculature endothelial cells.

Skachkov I., Luan Y., van Tiel S.T., van der Steen A.F., de Jong N., Bernsen M.R, & Kooiman K. (2018). SPIO labeling of endothelial cells using ultrasound and targeted microbubbles at diagnostic pressures. PLoS ONE, 13(9), e0204354.

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