All procedures involving mice were approved by the Institutional Animal Care and Use Committee (IACUC) of UT Southwestern Medical Center. Four-week-old female NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) purchased from the Jackson Laboratories were used in xenograft experiments. HCT116 or its clonal derivative cells (1.0 × 106) were injected subcutaneously into the left and right flanks of the mice. Drug treatment by intraperitoneal (i.p.) injection was started on the 17th day after tumor inoculation. In three cases, the matched duplicates showed >5-fold variation prior to drug treatment and were therefore excluded from the analysis. One group of mice received 20 mg/kg TG in 13 PBS by i.p. injection every day, and the other group received 13 PBS as a control. Tumor volume was estimated using digital calipers every 3 days and calculated with the formula (length × width2)/2. All mice were euthanized on the day 24 post drug treatment, and the tumors were resected and the tumor weight was recorded. Each of the tumors was homogenized in Buffer RLT of RNeasy mini kit (QIAGEN) using Precellys homogenizer (Bertin Instruments) at 4°C. RNA or proteins were isolated from the supernatant of each ruptured tumors after centrifugation with high speed using TRI re-agent following the manufacturer's protocols.
Il2rgtm1wjl szj nsg
Manufactured by Jackson ImmunoResearch
The Il2rgtm1Wjl/SzJ (NSG) is a laboratory mouse strain characterized by a targeted mutation in the interleukin-2 receptor gamma chain (Il2rg) gene. This mutation results in a deficiency in T cells, B cells, and natural killer cells, making the strain useful for engraftment of human cells and tissues.
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4 protocols using il2rgtm1wjl szj nsg
1
Xenograft Mouse Model for HCT116 Tumor
2
Adipocyte-Mediated Tumor Growth and Initiation
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All animal procedures were done using protocols approved by the University of Kentucky Animal Care and Use Committee. Six to eight week-old NOD.Cg-Prkdcscid IL2rgtm1Wjl/SzJ (NSG, The Jackson Laboratory) mice were used. Both male and female mice of equal numbers were included in each group. Mice were housed in barrier rooms with 12-h light/dark cycle. For tumor growth assay, control and CPT1A knockdown SW480 cells in 5% Matrigel suspension (5 × 105 cells in 100 µl) were mixed with 50 µl of freshly isolated human adipocytes (contain ~100,000 adipocytes) or PBS and inoculated subcutaneously. The tumor size was measured every week with a caliper, and the tumor volume was defined as (longest diameter) × (shortest diameter)2/2. At the end of experiments, tumors were harvested and subjected to mRNA and protein analysis. For tumor initiation assay, 100 or 1000 SW480 cells were mixed with Matrigel and adipocytes as described above and injected subcutaneously. The number of tumors formed was determined 3 months post injection.
3
Xenograft Mouse Model for HCT116 Tumor
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All procedures involving mice were approved by the Institutional Animal Care and Use Committee (IACUC) of UT Southwestern Medical Center. Four-week-old female NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) purchased from the Jackson Laboratories were used in xenograft experiments. HCT116 or its clonal derivative cells (1.0 × 106) were injected subcutaneously into the left and right flanks of the mice. Drug treatment by intraperitoneal (i.p.) injection was started on the 17th day after tumor inoculation. In three cases, the matched duplicates showed >5-fold variation prior to drug treatment and were therefore excluded from the analysis. One group of mice received 20 mg/kg TG in 13 PBS by i.p. injection every day, and the other group received 13 PBS as a control. Tumor volume was estimated using digital calipers every 3 days and calculated with the formula (length × width2)/2. All mice were euthanized on the day 24 post drug treatment, and the tumors were resected and the tumor weight was recorded. Each of the tumors was homogenized in Buffer RLT of RNeasy mini kit (QIAGEN) using Precellys homogenizer (Bertin Instruments) at 4°C. RNA or proteins were isolated from the supernatant of each ruptured tumors after centrifugation with high speed using TRI re-agent following the manufacturer's protocols.
4
Engrafting Patient-Derived AML/UCB Cells
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6–8 week-old NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG, Jackson Laboratories) mice were myeloablated with busulfan (30 mg/kg IP) at day 0. CD3-depleted primary AML patient samples or lineage-depleted UCB cells were ex vivo treated in the indicated conditions, and 4–34 × 106 (AML) or 12–14 × 104 (lin− UCB) cells were injected IV and left 8–10 weeks untreated. Engraftment was determined as the percentage of live human CD45-expressing cells in bone marrow as assessed by flow cytometry. Mice were randomized and blind-coded at the beginning of the experiment.
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