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Gammacell 40 exactor

Manufactured by Best Theratronics
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Sourced in Canada, United States

The Gammacell 40 Exactor is a laboratory irradiator designed for research applications. It utilizes a Cesium-137 sealed source to generate a gamma radiation field for sample exposure. The Gammacell 40 Exactor provides a uniform radiation field within the irradiation chamber, allowing for precise and consistent sample treatment.

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Lab products found in correlation

Lipofectamine 3000 reagent, by Thermo Fisher Scientific (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) X rad 320, by Precision X-Ray (3 mentions) Nvp lde225, by Selleck Chemicals (2 mentions) Crystal violet, by Merck Group (2 mentions) Pgc 1α sirna, by Santa Cruz Biotechnology (2 mentions) P53 sirna, by Santa Cruz Biotechnology (2 mentions) Celltiter glo reagent, by Promega (2 mentions) Synergy htx, by Agilent Technologies (2 mentions) White walled 96 well plate, by Corning (2 mentions) Apotome, by Zeiss (2 mentions) Hpaf 2, by American Type Culture Collection (2 mentions) Mia paca 2, by American Type Culture Collection (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Parafilm, by Amcor (1 mentions) Onalespib, by Selleck Chemicals (1 mentions) β cyclodextrin, by Merck Group (1 mentions) Vanox ahbt3, by Olympus (1 mentions) Perfection v500 photo, by Epson (1 mentions)

99 protocols using gammacell 40 exactor

1

Onalespib Drug Formulation and Irradiation

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Drug formulation for in vitro studies: 5 mg Onalespib (AT13387, Selleckchem) was dissolved in 200 µL DMSO and further diluted in cell culture medium before use. For in vivo studies: 5 mg Onalespib was dissolved in 200 µL DMSO and further diluted in 17. 5% β-Cyclodextrin (Sigma-Aldrich).
Irradiation: In vitro and in vivo experiments were irradiated with 137-Cs γ-rays using a Gammacell 40 Exactor (Best Theratronics Gammacell 40 Exactor, dose rate 1 Gy/min). For in vivo studies: Local irradiation of tumor xenografts (posterior leg) was achieved with specific collimators (Best Theratronics) resulting in a 3-cm radiation field.
Spiegelberg D., Abramenkovs A., Mortensen A.C., Lundsten S., Nestor M, & Stenerlöw B. (2020). The HSP90 inhibitor Onalespib exerts synergistic anti-cancer effects when combined with radiotherapy: an in vitro and in vivo approach. Scientific Reports, 10, 5923.
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2

Quantifying Radiation-Induced Cell Death

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Cell death in response to irradiations was assayed by measuring colony formation with and without increasing doses of gamma irradiation (0, 2, 4 and 6 Gy) treatment using a Gammacell® 40 Exactor (Best Theratronics Ltd, Kanata, ON, CA). 300 cells per well were seeded on a 6-well plate and incubated for 8 h before being treated with increasing dose of gamma radiation (2, 4 and 6 Gy) using a Gammacell® 40 Exactor (Best Theratronics Ltd, Kanata, ON, CA). Culture medium was replaced every three days, and cells were incubated for 10–11 days or until surviving individual cells at T0 had divided sufficiently to form a visible colony (estimated to be 60 cells). Cells were fixed with methanol-acetic acid (3:1) and stained with 0.4% crystal violet. Excess dye was removed by immersing the plate in clean water twice. The 6-well plates were imaged using a chemidoc (Bio-Rad, Hercules, CA, USA) and the number of colonies was counted in CellProfiler using an automated analysis pipeline. Relative colony formation (%) as measured by the number of colonies from treated well divided by the number of colonies in the untreated control and plotted.
Tropée R., de la Peña Avalos B., Gough M., Snell C., Duijf P.H, & Dray E. (2020). The SWI/SNF subunit SMARCD3 regulates cell cycle progression and predicts survival outcome in ER+ breast cancer. Breast cancer research and treatment, 185(3), 601-614.
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3

Cell Culture and Clonogenic Assay

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U2OS, HEK293T and HeLa cell lines were cultured in DMEM medium with 10% fetal bovine serum (Sigma), 1X penicillin/streptomycin (Gibco). All cells were cultured in an atmosphere of 5% CO2 at 37°C. Lentivirus-mediated shRNA knockdown and siRNA knockdown were performed as reported (Lu et al., 2014a (link)). The sequences of siRNA and shRNA are listed in Table S2. Polyplus JetPrime® was used for DNA transfection. γ rays were generated using a cesium-137 source (Gammacell Exactor 40, Best Theratronics). Radiation dose is 10 Gy, and post-irradiation recovery time is indicated in the figure legends. For the colony formation assay, cells were irradiated and then stained with 2% methylene blue in 5% ethanol 10 days after IR. The colonies with over 50 cells were counted. The results are presented as mean ± SEM from three independent experiments with P value by Student’s t-test.
Lu H., Shamanna R.A., Keijzers G., Anand R., Rasmussen L.J., Cejka P., Croteau D.L, & Bohr V.A. (2016). RECQL4 promotes DNA end resection in repair of DNA double-strand breaks. Cell reports, 16(1), 161-173.
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4

Multifractional Gamma Radiation Exposure

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Cells were exposed repeatedly to gamma radiation from a 137Cs source Gamma Cell® Exactor 40 (Best Theratronics, Ottawa, ON, Canada) at room temperature, at 0.77 Gy/min. Non-irradiated control samples were sham exposed. The radiation exposure of VH10 fibroblasts always started on a Monday, followed by exposures on Wednesday and Friday (3 fractions per week); VH10 fibroblasts were irradiated for a total of 3 weeks (21 days), and passaged once a week. Multifractional radiation exposure of AHH-1 cells always started on a Monday and was followed by exposures on Tuesday–Friday (5 fractions per week), AHH-1 cells were irradiated for two weeks, and passaged three times a week (Figure 1).
Akuwudike P., Tartas A., López-Riego M., Toma-Dasu I., Wojcik A, & Lundholm L. (2022). Cell Type-Specific Patterns in the Accumulation of DNA Damage Following Multifractional Radiation Exposure. International Journal of Molecular Sciences, 23(21), 12861.
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5

Adherent Cell Culture Irradiation

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Cells were cultured in adherent monolayer and irradiated in a Gammacell 40 Exactor (Best Theratronics) or sham irradiated. When removed from the incubator, culture plates were sealed with Parafilm (Bemis) until returned.
Robertson F.L., O'Duibhir E., Gangoso E., Bressan R.B., Bulstrode H., Marqués-Torrejón M.Á., Ferguson K.M., Blin C., Grant V., Alfazema N., Morrison G.M, & Pollard S.M. (2023). Elevated FOXG1 in glioblastoma stem cells cooperates with Wnt/β-catenin to induce exit from quiescence. Cell reports, 42(6).
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6

Metabolomic Analysis of Irradiated Cells

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Cell culture flasks containing 4 × 106 cells were γ-irradiated in groups of six using a Gammacell 40 Exactor (Best Theratronics, Ottawa, Canada). The irradiator was fitted with two 137Cs sources (above and below) with an activity of 1800 Ci/67 TBq and delivering 1.0 Gy/min. Nominal radiation doses of 0, 1, 2 and 4 Gy were employed. Sham irradiation (0 Gy) was achieved by placing the culture flasks in the irradiator for 2 min without irradiation. Immediately after irradiation, cells were cultured for a further 4 h. One flask of HepG2 cells at each dose was used for FACS and fluorescence immunohistochemical (IHC) analyses and five flasks were used for metabolomic analysis. HMCL-7304 myotubes were subjected only to metabolomic analysis after irradiation.
Wang M., Keogh A., Treves S., Idle J.R, & Beyoğlu D. (2016). The metabolomic profile of gamma-irradiated human hepatoma and muscle cells reveals metabolic changes consistent with the Warburg effect. PeerJ, 4, e1624.
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7

Dosimetry Calibration for Ir-192 Brachytherapy

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Since TLD-100 response varies significantly with radiation energy, to perform accurate dosimetry for BT treatments with Ir-192, calibration was performed indirectly by exposing the dosimeters to three different radiation fields:

250 kV X-ray spectrum with 1 mm Cu added filtration (HVL = 2.1 mm Cu) with a Siemens Stabilipan 2 X-ray tube (Eaverage = 114 keV obtained with software TASMICS [52 (link)]).

Cesium-137 (Cs-137) with Gammacell® 40 Exactor by Best Theratronics (Eaverage = 662 keV).

Cobalt-60 (Co-60) with the Gammacell® 220 provided by Atomic Energy of Canada Limited (Eaverage = 1250 keV).

Ir-192 was not used for calibration because accurate dose delivery in positions close to the source is made challenging by steep dose gradients [53 (link),54 ].
The batch of 20 TLDs was randomly divided into four groups of 5 dosimeters. One group was used for radiation background, while the remaining groups were irradiated with known radiation doses of 0.5, 1.5, and 5 Gy, with each one being exposed to the three radiation fields. For each exposure, a graph of reading vs dose was plotted and, through a linear fit the calibration factor (CF), was evaluated as the slope of the line.
A graph of CF vs energy was then plotted and the factor for Ir-192 was obtained as linear interpolation [55 (link),56 (link),57 (link)].
Manna F., Pugliese M., Buonanno F., Gherardi F., Iannacone E., La Verde G., Muto P, & Arrichiello C. (2023). Use of Thermoluminescence Dosimetry for QA in High-Dose-Rate Skin Surface Brachytherapy with Custom-Flap Applicator. Sensors (Basel, Switzerland), 23(7), 3592.
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8

Precision Gamma Irradiation of Mice

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For all RT, a dual source 137Cs Gammacell 40 Exactor (Best Theratronics) was used with a custom-built lead shield comprising two 4 cm thick lead slabs, a lucite immobilization platform, and a metal frame for positioning within the Gammacell center RT field (fig. S2). 3 cm corner posts separated the two slabs from each other to accommodate the lucite platform, and sides did not require shielding given the photon source directionality. A 2 cm aperture was cut through each lead slab to allow local irradiation. Prior to use, dosimetric studies using standard film-based and thermoluminescent dosimeter techniques showed the average dose in the center of the aperture to be 68.5% ± 3% (mean ± S.D.) of that delivered with a fully open field. Dose at 2 cm inferior to the aperture was 95.6% lower than in the aperture center. The Gammacell holds a published dose rate accuracy of ± 15%. Immediately prior to RT, mice were anesthetized via 87.5 mg/kg ketamine and 12.5 mg/kg xylazine intraperitoneally (i.p.), immobilized on the lucite platform with surgical tape, and irradiated individually according to the calibrated Gammacell dose rate of 0.6 Gy min−1.
Miller M.A., Chandra R., Cuccarese M.F., Pfirschke C., Engblom C., Stapleton S., Adhikary U., Kohler R.H., Pittet M.J, & Weissleder R. (2017). Radiation therapy primes tumors for nanotherapeutic delivery via macrophage-mediated vascular bursts.. Science translational medicine, 9(392), eaal0225.
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9

Tumor Cell Proliferation Assays

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To measure effects of inhibitors in combination with radiation, tumor cells were plated in 96-well plates at a density of 0.2 × 106 cells per well and cultured in the presence of DMSO, 50 nM YM155, or 10 ug/ml S12. After 24hrs, cells were subjected to 0, 0.25, or 0.5 Gy radiation using a Gammacell 40 Exactor (Low-dose cesium 137 irradiator, Best Theratronics Ltd., Ottawa, Ontario, Canada). Cells were cultured for an additional 24 hr, and [methyl-3H]thymidine assays were performed as described above.
To measure effects of inhibitors in combination with the SHH antagonist NVP-LDE225 (Selleck Chemicals, S2151), tumor cells were plated in 96 well plates at 0.2 × 106 cells/well and cultured with increasing doses of LDE225 or a single dose of Survivin antagonist (10μg/ml S12, 20nM YM255) alone or in combination with LDE225 as indicated. Cells were cultured for 48hrs and [methyl-3H]thymidine assays were performed as described above.
Brun S.N., Markant S.L., Esparza L.A., Garcia G., Terry D., Huang J.M., Pavlyukov M.S., Li X.N., Grant G.A., Crawford J.R., Levy M.L., Conway E.M., Smith L.H., Nakano I., Berezov A., Greene M.I., Wang Q, & Wechsler-Reya R.J. (2014). Survivin as a therapeutic target in Sonic hedgehog-driven medulloblastoma. Oncogene, 34(29), 3770-3779.
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10

Tumor Cell Proliferation Assays

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To measure effects of inhibitors in combination with radiation, tumor cells were plated in 96-well plates at a density of 0.2 × 106 cells per well and cultured in the presence of DMSO, 50 nM YM155, or 10 ug/ml S12. After 24hrs, cells were subjected to 0, 0.25, or 0.5 Gy radiation using a Gammacell 40 Exactor (Low-dose cesium 137 irradiator, Best Theratronics Ltd., Ottawa, Ontario, Canada). Cells were cultured for an additional 24 hr, and [methyl-3H]thymidine assays were performed as described above.
To measure effects of inhibitors in combination with the SHH antagonist NVP-LDE225 (Selleck Chemicals, S2151), tumor cells were plated in 96 well plates at 0.2 × 106 cells/well and cultured with increasing doses of LDE225 or a single dose of Survivin antagonist (10μg/ml S12, 20nM YM255) alone or in combination with LDE225 as indicated. Cells were cultured for 48hrs and [methyl-3H]thymidine assays were performed as described above.
Brun S.N., Markant S.L., Esparza L.A., Garcia G., Terry D., Huang J.M., Pavlyukov M.S., Li X.N., Grant G.A., Crawford J.R., Levy M.L., Conway E.M., Smith L.H., Nakano I., Berezov A., Greene M.I., Wang Q, & Wechsler-Reya R.J. (2014). Survivin as a therapeutic target in Sonic hedgehog-driven medulloblastoma. Oncogene, 34(29), 3770-3779.
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