T 25 culture flasks

Manufactured by BD

Sourced in United States

The T-25 culture flask is a laboratory equipment used for cell culture applications. It is a small, rectangular-shaped flask with a surface area of approximately 25 square centimeters, designed to provide a controlled environment for the growth and maintenance of cells in vitro.

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Lab products found in correlation

Truebeam, by Agilent Technologies (3 mentions) Paclitaxel, by Merck Group (2 mentions) Retronectin, by Takara Bio (1 mentions) Neurocult ns a differentiation kit, by STEMCELL (1 mentions) Basic fibroblast growth factor (bfgf), by BD (1 mentions) Epidermal growth factor (egf), by BD (1 mentions) Igf 1, by BD (1 mentions) Neurocult ns a proliferation kit, by STEMCELL (1 mentions) Igf 1, by Merck Group (1 mentions) Basic fibroblast growth factor (bfgf), by Merck Group (1 mentions) L 15 medium, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) 96 well tissue culture plate, by Corning (1 mentions) 6 well tissue culture plate, by Corning (1 mentions) 35 mm petri dishes, by BD (1 mentions) Accutase, by Innovative Cell Technologies (1 mentions)

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9 protocols using t 25 culture flasks

Retroviral Transduction of PBMCs

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For retroviral transduction, PBMCs were stimulated with immobilized anti-CD3 antibody OKT-3 (eBiosciences, San Diego, CA) and RetroNectin (Takara Bio) for 4 days. Then, we prepared three groups of retroviral mixtures including RV-CAR only, RV-iReporter only, and a 1:1 mixture of RV-CAR and RV-iReporter. These mixtures were applied to vector-preloaded RetroNectin-coated 24-well plates and centrifuged at 2,000 × g for 2 hr at 32°C. Then, pre-stimulated PBMCs were added to the preloaded plates and centrifuged at 1,000 × g for 10 min at 32°C. Cells were cultured at 37°C for 5 hr and then transferred to T-25 culture flasks (BD Falcon).

Uchibori R., Teruya T., Ido H., Ohmine K., Sehara Y., Urabe M., Mizukami H., Mineno J, & Ozawa K. (2018). Functional Analysis of an Inducible Promoter Driven by Activation Signals from a Chimeric Antigen Receptor. Molecular Therapy Oncolytics, 12, 16-25.

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Induction of ESSC Proliferation and Differentiation

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E18 ESSCs were cultured using the NeuroCult NS-A Proliferation Kit and NeuroCult NS-A Differentiation Kit (rat) from Stem Cell Technologies, for the induction of proliferation and differentiation, respectively. The growth factor supplements consisting of insulin-like growth factor-1 (IGF-1, Sigma I 8779), leukaemia inhibitory factor (LIF, Sigma L 5158), epidermal growth factor (EGF, Sigma E 4127) and basic fibroblast growth factor (bFGF, Sigma F 0291) were purchased from Sigma. The concentrations of these growth factors used were 20 ng/mL for the EGF and bFGF, 100 ng/mL for the IGF-1 and 20 ng/mL for the LIF, in accordance with our recently published report [7 (link)]. The ESSCs were plated in T-25 culture flasks (BD Falcon) in triplicate, under five different conditions (Figure 1): group A (no growth factor); group B (EGF + bFGF); group C (EGF + bFGF + LIF); group D (EGF + bFGF + IGF-1); and group E (EGF + bFGF + LIF + IGF-1).

Pati S., Supeno N.E., Muthuraju S., Abdul Hadi R., Ghani A.R., Idris F.M., Maletic-Savatic M., Abdullah J.M, & Jaafar H. (2014). MicroRNA Profiling Reveals Unique miRNA Signatures in IGF-1 Treated Embryonic Striatal Stem Cell Fate Decisions in Striatal Neurogenesis In Vitro. BioMed Research International, 2014, 503162.

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Culture of C6/36 Aedes albopictus Cells

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C6/36 (Aedes albopictus) cells were grown and maintained in L15 medium (Gibco, Grand Island, NY) supplemented with 10% FBS (Gibco), 1% L-glutamine (Gibco) and 1% penicillin-streptomycin (Gibco) at 27°C. The cells were grown as a monolayer in T-25 culture flasks (BD Falcon, Franklin Lakes, NJ), 6-well tissue culture plates, or 96-well tissue culture plates (Corning, Tewksbury, MA).

Lee S.B., Chen J., Aimanova K.G, & Gill S.S. (2014). Aedes cadherin mediates the in vivo toxicity of the Cry11Aa toxin to Aedes aegypti. Peptides, 68, 140-147.

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Synergistic Radiation-Paclitaxel Treatment

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Cells were seeded (2× 105) in T‐25 culture flasks (BD‐Falcon), allowed to adhere overnight, and irradiated with 10 MV x‐rays at dose rates of 2400 MU/min or 400 MU/min by using TrueBeam (Varian Medical Systems, CA) with 10×‐FFF mode. The radiation dose was administered to melanoma and normal cells as shown in Figure S1 and Table S1. Each group's duplicate set of cells was pretreated with Paclitaxel (50 nM in DMSO; Sigma Aldrich) for 2 h prior to each irradiating step (Rad ×1, Rad ×2, Rad ×3, and Rad ×4). The cell culture medium was changed after 2 h of each radiation. Cells were treated with Paclitaxel only 2 h prior to each irradiation. The titrated dose of 50 nM Paclitaxel was not toxic to normal skin cells for the entire duration of the experiment. After 24 h of treatment, cells were plated. Delaying the plating of cells after irradiating allows cells a more accurate concentration versus time exposure.

Lodhi N., Nagpal P., Sarojini S., Keck M., Chiu Y.M., Parvez Z., Adrianzen L, & Suh K.S. (2022). Synergetic effect of high dose rate radiations (10× FFF/2400 MU/min/10 MV x‐rays) and paclitaxel selectively eliminates melanoma cells. Cancer Reports, 6(2), e1733.

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Irradiation of Cancer Cells

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Cells were seeded (5×10⁵) in T-25 culture flasks (BD-Falcon; BD-Biosciences, Durham, North Carolina, USA), allowed to adhere overnight, and irradiated with 10 MV X-rays at dose rates of 400 or 2400 MU/min using TrueBeam (Varian Medical Systems, Palo Alto, California, USA) with 10X FFF. The total dose range of 10 MV X-rays was 0.25–8 Gy (100–1000 Gy data not shown) for multiple experiments.

Sarojini S., Pecora A., Milinovikj N., Barbiere J., Gupta S., Hussain Z.M., Tuna M., Jiang J., Adrianzen L., Jun J., Catello L., Sanchez D., Agarwal N., Jeong S., Jin Y., Remache Y., Goy A., Ndlovu A., Ingenito A, & Suh K.S. (2015). A combination of high dose rate (10X FFF/2400 MU/min/10 MV X-rays) and total low dose (0.5 Gy) induces a higher rate of apoptosis in melanoma cells in vitro and superior preservation of normal melanocytes. Melanoma Research, 25(5), 376-389.

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Culturing Human Cell Lines for Research

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The human cell lines K-562 (CML), THP-1 (acute myeloid leukemia), HEK-293 (human embryonic kidney) were obtained from the NCCS, Pune, India. Whereas, peripheral blood mononuclear cells (PBMCs) were procured from HiMedia (Mumbai, India). K-562, THP-1 and PBMCs were cultured in RPMI-1640 medium and HEK-293 cells in DMEM high glucose, supplemented with 10% FBS, 2 mM glutamine, 1% penicillin and streptomycin, and kept at 37°C in a humidified incubator with 5% CO₂. Stock cultures were maintained in the exponential growth phase by passaging with their respective growth medium after 2–3 days in T-25 culture flasks (BD Falcon, U.S.A.).

Rai Y., Yadav P., Kumari N., Kalra N, & Bhatt A.N. (2019). Hexokinase II inhibition by 3-bromopyruvate sensitizes myeloid leukemic cells K-562 to anti-leukemic drug, daunorubicin. Bioscience Reports, 39(9), BSR20190880.

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Cell Isolation and Culture Protocol

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Equipment consisted of scalpels, 100-µm nylon cell strainer, T25 culture flasks, 35-mm Petri dishes, 6- and 24-well culture plates (all from BD Falcon), and 6- and 24-well culture inserts (cat Nos. PIHT 30R48 and PIHT 12R 48, respectively; Millicell).

Nanduri V., Tattikota S.M., T. A.R., Sriramagiri V.R., Kantipudi S, & Pande G. (2014). Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes. Orthopaedic Journal of Sports Medicine, 2(6), 2325967114539122.

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Astrocyte Isolation from Ferret Cortex

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The culture procedure is modified from the method described by McCarthy and de Vellis (McCarthy & de Vellis 1980 (link)). Briefly, ferrets at P31 were sacrificed and the occipital portion of their cortices immediately dissected, cleaned from its meninges and put in a petri dish containing Hank’s Balanced Salt Solution (HBSS). The tissue block was minced and exposed to 0.05% Trypsin–EDTA solution for 10-20 min. The tissue pieces were then mechanically triturated and serum medium (DMEM/F12 + 10% dialyzed fetal bovine serum) was added to inactivate trypsin. The cell suspension was centrifuged at 1000rpm for 3min, resuspended in SILAC growth medium and plated in T-25 culture flasks (Falcon, BD). The flasks were kept in an incubator at 37°C and 5% CO₂. Growth media were changed every 2-3 days. Between culture days 7 and 10 (C7-10), flasks were agitated on an orbital shaker (250 rpm, 37°C, 12–18h) in order to remove cells other than astrocytes, such as oligodendrocytes progenitors and microglia. An astrocyte purity of ~98% was achieved after supernatant removal (McCarthy & de Vellis 1980 (link)). Before reaching confluence, at around Cultured day (C)12-15, cells were detached with Accutase (Innovative Cell Technologies) and passed to T-75 flasks.

Trindade P., Hampton B., Manhães A, & Medina A. (2016). Developmental alcohol exposure leads to a persistent change on astrocyte secretome. Journal of neurochemistry, 137(5), 730-743.

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Combinatorial Effects of Paclitaxel and Radiotherapy

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Cells were seeded (2x 105) in T-25 culture flasks (BD-Falcon), allowed to adhere overnight, and irradiated with 10 MV x-rays at dose rates of 2400 MU/min or 400MU/min by using TrueBeam (Varian Medical Systems, CA) with 10X-FFF mode. The radiation dose was administered to melanoma and normal cells as shown in Supplementary Figure 1 and Supplementary Table 1. Where a duplicate set of cells of each group was pretreated with paclitaxel (50nM in DMSO; Sigma Aldrich,A) for two hours prior to each irradiating step (Rad x1, Rad x2, Rad x3, and Rad x4). The cell culture medium was changed after two hours of each radiation. Cells were treated with paclitaxel only two hours prior to each irradiation. The titrated dose of 50nM paclitaxel was not toxic to normal skin cells for the entire duration of the experiment.

Lodhi N., Sarojini S., Keck M.K., Nagpal P., Chiu Y.M., Parvez Z., Adrianzen L., & Suh K.S. (2021). Combination of High Dose Rate Radiations (10X FFF/2400 MU/min/10 MV X-rays) and Paclitaxel Selectively Eliminates Melanoma Cells.

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