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T25 tissue culture flasks

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The T25 tissue culture flask is a laboratory equipment item used for the in vitro cultivation of cells. It provides a sterile, controlled environment for the growth and maintenance of cell cultures. The flask has a surface area of 25 cm2 and is made of clear, durable plastic material.

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Lab products found in correlation

Collagenase type 2, by Worthington (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Trypsin, by Thermo Fisher Scientific (2 mentions) Gentamicin, by Thermo Fisher Scientific (2 mentions) Laminin, by Merck Group (2 mentions) Advanced dmem medium, by Thermo Fisher Scientific (2 mentions) F98 rat glioma cells, by American Type Culture Collection (2 mentions) α mem, by Thermo Fisher Scientific (2 mentions) α mem medium, by Thermo Fisher Scientific (1 mentions) Bleomycin blm, by Merck Group (1 mentions) Doxorubicin hydrochloride dox, by Merck Group (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Gentamycin, by Thermo Fisher Scientific (1 mentions) Murine laminin, by Merck Group (1 mentions)

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T25 flasks (45 citations) Culture flasks (15 citations)

6 protocols using t25 tissue culture flasks

1

Isolation of Primary Murine Vascular Smooth Muscle Cells

Cited 3 times
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Mice were euthanized by cervical dislocation. Primary murine VSMCs were isolated as previously described25 (link)33 (link)34 (link). After removal of adventitia, the aorta was cut open to expose the endothelial layer under a dissection microscope. Tissues from eight animals were pooled and incubated with 1 mg ml−1 trypsin (Invitrogen) for 10 min in order to remove any remaining adventitia and endothelium. After a further overnight incubation at 37 °C in a humidified atmosphere of 95% air/5% CO2 in α-MEM medium (Invitrogen) supplemented with 10% Fetal Bovine Serum (Invitrogen) and 1% gentamicin (Invitrogen), tissues were digested with 425 U/ml collagenase type II (Worthington Biochemical Corporation, Lakewood, USA) for 5 hrs. Medial cells were released and cell suspensions were centrifuged at 2000 g for 5 min. The cell pellet was washed and resuspended in the above mentioned culture medium. Isolated VSMCs were cultured with growth medium for two passages in T25 tissue culture flasks (Greiner Bio-one, GmbH, Frickenhausen, Baden-Wurttemberg, Germany) coated with 0.25 μg/cm2 laminin (Sigma) to promote maintenance of the contractile differentiation state47 (link).
Zhu D., Hadoke P.W., Wu J., Vesey A.T., Lerman D.A., Dweck M.R., Newby D.E., Smith L.B, & MacRae V.E. (2016). Ablation of the androgen receptor from vascular smooth muscle cells demonstrates a role for testosterone in vascular calcification. Scientific Reports, 6, 24807.
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2

Culturing F98 Rat Glioma Cells

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The F98 rat glioma cells were obtained from the American Type Culture Collection (Manassas, VA). The cell line was maintained in Advanced DMEM medium (Thermo Fisher Scientific, Carlsbad, CA) supplemented with 2% heat-inactivated fetal bovine serum (FBS), 25 mM HEPES buffer, 100 U/mL penicillin and 100 μg/mL streptomycin at 37°C, 5% CO2 and 95% humidity. The F98 cells were grown as monolayers in T-25 tissue culture flasks, (Greiner BioOne Frickenhausen Germany). Bleomycin (BLM) was obtained from Sigma Aldrich (St. Louis, MO).
Shin D., Nguyen L., Le M.T., Ju D., Le J.N., Berg K, & Hirschberg H. (2019). The Effects of Low Irradiance Long Duration Photochemical Internalization on Glioma Spheroids. Photodiagnosis and photodynamic therapy, 26, 442-447.
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3

Expansion of Bone Marrow-Derived MNCs

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MNCs from unfractionated BM aspirates (see above for isolation) were seeded in T25 tissue culture flasks (Greiner Bio-One, Monroe NC) at 107/7 mL of EGM-2 Endothelial Growth Media. Confluence was noted at day 21. At this time, the adherent cells were serially passaged.
Moore C.A., Siddiqui Z., Carney G.J., Naaldijk Y., Guiro K., Ferrer A.I., Sherman L.S., Guvendiren M., Kumar V.A, & Rameshwar P. (2021). A 3D Bioprinted Material That Recapitulates the Perivascular Bone Marrow Structure for Sustained Hematopoietic and Cancer Models. Polymers, 13(4), 480.
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4

Culturing Murine Macrophages and Rat Glioma Cells

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The murine macrophages RAW264.7 (designated Ma) and F98 rat glioma cells were obtained from the American Type Culture Collection (Manassas, VA). Both cell lines were maintained in Advanced DMEM medium (Thermo Fisher Scientific, Carlsbad, CA) supplemented with 2% heat-inactivated fetal bovine serum (FBS), 25mM HEPES buffer, 100 U/ml penicillin and 100 µg/ml streptomycin at 37 °C, 5% CO2 and 95% humidity. The F98 and Ma cells were grown as monolayers in T-25 tissue culture flasks, Greiner BioOne (Frickenhausen Germany) and in 9 cm flat-bottomed square dishes (Simport Scientific Beloeil, QC, Canada) respectively. Doxorubicin Hydrochloride (DOX) was obtained from Sigma Aldrich (St. Louis, MO).
Shin D., Christie C., Ju D., Nair R.K., Molina S., Berg K., Krasieva T.B., Madsen S.J, & Hirschberg H. (2017). Photochemical internalization enhanced macrophage delivered chemotherapy. Photodiagnosis and photodynamic therapy, 21, 156-162.
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5

Murine Vascular Smooth Muscle Cell Isolation

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Mice were euthanized by cervical dislocation. Primary murine VSMCs were isolated as described [24] (link). Briefly, after removal of the adventitia, the aorta was opened to expose the endothelial layer under a dissection microscope. Tissues from eight animals were pooled and incubated with 1 mg ml− 1 trypsin (Invitrogen, Paisley, UK) for 10 min in order to enable the removal of any remaining adventitia and endothelium through further dissection. Following overnight incubation at 37 °C in a humidified atmosphere of 95% air/5% CO2 in “growth medium” (α-MEM supplemented with 10% fetal bovine serum and 1% gentamicin, all from Invitrogen), tissues were digested with 425 U/ml collagenase type II (Worthington Biochemical Corporation, Lakewood, USA) for 5 h. Cell suspensions were centrifuged at 2000 g for 5 min. The cell pellet was washed and resuspended in growth medium. Isolated VSMCs were passaged in growth medium twice in T25 tissue culture flasks (Greiner Bio-one, GmbH, Frickenhausen, Baden-Wurttemberg, Germany) coated with 0.25 μg/cm2 laminin (Sigma, Poole, UK) to promote maintenance of the contractile differentiation state [17] (link). VSMCs were subsequently seeded at a density of 1.5 × 104/cm2 in 12-well plates.
Zhu D., Rashdan N.A., Chapman K.E., Hadoke P.W, & MacRae V.E. (2016). A novel role for the mineralocorticoid receptor in glucocorticoid driven vascular calcification. Vascular Pharmacology, 86, 87-93.
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6

Isolation and Culture of Primary Vascular Smooth Muscle Cells

Cited 2 times
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Primary VSMCs were isolated from 5-week-old WT male C57BL/6 mice. Following dissection of the aorta, the adventitia was detached and the blood vessel was opened to expose the endothelial layer of cells (Mackenzie et al. 2011 (link), Zhu et al. 2011 (link), 2013 (link)). Aortae isolated from 16 mice were then digested with 1 mg/ml trypsin for 10 min to remove any remaining endothelium and adventitia. This was followed by an overnight incubation at 37 °C in a humidified atmosphere of 95% air/5% CO2 in growth medium (α-MEM (Invitrogen) supplemented with 10% FCS (Invitrogen) and 1% gentamycin (Invitrogen)). Tissues were then digested in 425 U/ml collagenase type II for 5 h. Before experimental studies, isolated VSMCs were expanded in a growth medium for two passages in T25 tissue culture flasks (Greiner Bio-One GmbH, Frickenhausen, Baden-Württemberg, Germany) coated with 0.25 μg/cm2 murine laminin (Sigma) to prevent VSMC de-differentiation (Johnson et al. 2008 (link)).
Zhu D., Mackenzie N.C., Millan J.L., Farquharson C, & MacRae V.E. (2014). Upregulation of IGF2 expression during vascular calcification. Journal of molecular endocrinology, 52(2), 77-85.
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