The crypts were isolated from the small intestines of mice, and isolated crypts were cultured as previously reported [45 (link)]. The organoids were cultured onto four-well plates at the same ratio, and then exposed to 137Cs γ-rays from a Gamma Cell-3000 irradiator (MDS Nordion International, Kanata, Ontario, Canada). Metformin (20 μM; Sigma-Aldrich, St. Louis, MO, USA) was administered after irradiation on day 0, and the medium was changed every three days. For examination of intestinal organoid viability after irradiation, we performed methylthiazolyl tertrazolium (MTT) assays, as previously reported [46 (link)]. Briefly, organoids were incubated with 10% MTT for 2–3 h, and the insoluble formazan crystal were dissolved in solubilization solution. MTT-stained organoids indicated surviving cells.
Gammacell 3000
Manufactured by Nordion
Sourced in Canada
The Gammacell 3000 is a gamma irradiator designed for research applications. It uses a sealed radioactive source to provide a consistent and uniform irradiation field. The Gammacell 3000 is capable of delivering precise doses of gamma radiation to samples placed inside the irradiation chamber.
Automatically generated - may contain errors
Lab products found in correlation
Metformin, by Merck Group (1 mentions)
Auranofin, by Merck Group (1 mentions)
Tunicamycin, by Merck Group (1 mentions)
Tudca, by Tokyo Chemical Industry (1 mentions)
Bromodeoxyuridine (brdu), by Merck Group (1 mentions)
Quinpirole, by Bio-Techne (1 mentions)
Aripiprazole, by Bio-Techne (1 mentions)
Haloperidol, by Bio-Techne (1 mentions)
Thioridazine, by Cayman Chemical (1 mentions)
A 769662, by LC Laboratories (1 mentions)
Entinostat, by Merck Group (1 mentions)
Vorinostat, by Merck Group (1 mentions)
Hsc expansion cocktail, by Miltenyi Biotec (1 mentions)
Pkh67 dye, by Merck Group (1 mentions)
Clodronate liposome, by Liposoma (1 mentions)
12 protocols using gammacell 3000
1
Irradiation and Metformin Effect on Intestinal Organoids
2
Radiation-Induced Lung Injury in Mice
3
Ionizing Radiation Impacts on Embryonic and Postnatal Liver
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Pregnant mice were subjected to 4–6 Gy of IR (Gammacell 3000, MDS Nordion, Germany) at defined embryonic stages. At 0, 6, 16, and 24 hours after IR, pregnant mice were sacrificed, and embryonic livers were dissected for cell cycle analysis and other experiments. P0 to P56 mice were also subjected to 2 Gy of IR, and the liver cells were isolated at multiple time points.
4
Gamma-ray Irradiation Protocol
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Auranofin and tunicamycin were purchased from Sigma-Aldrich (St Louis, MO, United States). TUDCA was purchased from TCI America (Portland, OR, United States). Irradiation was performed at room temperature using a 137Cs gamma-ray source, Gammacell 3000, manufactured by Nordion (Ottawa, ON, Canada).
5
Tumor Cell Irradiation Protocol
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Tumor cells were irradiated at 15 000 rads using Gammacell 3000 (MDS Nordion, Canada).
6
Improving Survival After Irradiation
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For irradiation, 10-week-old male C57BL/6 J mice were exposed to single total body irradiation of 1100 Rad using a Gamma cell 3000 (Nordion Inc., Ottawa, Canada) at the KAIST Laboratory Animal Resource Center (Daejeon, Rep. of Korea). After irradiation, mice were treated orally with vehicle or Amuc_1409* (9 μg per mouse) daily for 8 days for survival analysis (Figs. 4a ) and for 5 days for intestinal tissue isolation (Fig. 4c ). For BrdU staining, mice were injected intraperitoneally 6 h before euthanasia with 200 μL of BrdU (Sigma-Aldrich) dissolved in PBS to 5 mg/mL, and intestinal tissue samples were collected at day 5 post-irradiation.
7
Whole-body Irradiation Dosimetry Protocol
8
Fluorescent Visualization of Hematopoietic Stem Cell Transplantation
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To fluorescently visualize the donor bone marrow cells, the β‐actin‐DsRed transgenic mouse (20–35 g body weight; 12–25 weeks old, C57BL/6), and the wildtype C57BL/6 mouse (20–35 g; 12–25 weeks, C57BL/6) were used as donors and recipients, respectively, of hematopoietic stem cell transplantation. At 7–12 h before HSPC infusion, recipients were sub‐lethally irradiated with 6–9.5 Gy using a gamma irradiator (Gamma cell 3000, Nordion Inc.). 3–4 × 104 purified HSPCs were systemically transferred to each recipient mouse via tail‐vein injection. From representative three HSCT recipient mice for performing in vivo imaging experiments, bone marrow reconstitution was successfully assessed at 4, 8, and 12 weeks post‐transplantation (Figure S1 ).
9
Pharmacological Treatments and Irradiation
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Aripiprazole, quinpirole, and haloperidol were purchased from Tocris (Ellisville, MO, USA). A769662 was obtained from LC Laboratories (Woburn, MA, USA). Thioridazine was obtained from Cayman Chemical (Ann Arbor, MI, USA). Irradiation was performed at room temperature by using a 137Cs gamma‐ray source Gammacell 3000 manufactured by Nordion (Ottawa, ON, Canada).
10
Assessing Efficacy of HDAC Inhibitors
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For irradiation, a Cs-137 source (Gammacell 3000; Nordion, Ottawa, ON, Canada) was used. Vorinostat and entinostat were obtained from Sigma Aldrich GmbH (Taufkirchen, Germany). The compound library of hydroxamic acid- and benzamide-type HDACi derivatives was provided by T. Kurz and F. K. Hansen (Institute of Pharmaceutical and Medicinal Chemistry, Heinrich Heine University Duesseldorf, Düsseldorf, Germany). Synthesis and biological activities of the HDAC inhibitory compounds KSK64 [30 (link)] and DDK137 [79 (link)] were already described. More detailed information regarding the synthesis of the various HDACi is also provided by Pflieger et al. [80 (link)], Mackwitz et al. [81 (link)], and Krieger et al. [82 (link)]. Chemical structures of the HDACi tested are summarized in Supplementary Figure S1 . HDAC inhibitory activity of the compounds was confirmed by analyses of hyperacetylation of histones as concluded from enzyme assays and cellular HDAC pan assays [20 (link),30 (link),33 (link),79 (link),82 (link)].
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