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18 protocols using nod cg prkdcscid il2rgtm1sug jictac

1

NOG Mouse Bladder Organoid Implantation

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All experiments using animals were performed according to protocols approved by the Institutional Animal Care and Use Committee (IACUC) at Columbia University Medical Center. 6–8 week old male NOG (NOD.Cg-PrkdcscidIl2rgtm1Sug/JicTac; Taconic) mice were used for ultrasound-guided delivery of organoids diluted in 50% Matrigel/media into the mouse bladder wall. Mice were observed for clinical signs of disease daily, and mice were euthanized according to IACUC-approved criteria.
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2

Adoptive T-cell Transfer in Melanoma Xenograft Model

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Immune deficient NOG mice (NOD.Cg-PrkdcScidIL2rgtm1sug/JicTac, Taconic) were housed in individually ventilated cages at University of Copenhagen, and supplied only autoclaved or sterilized water, food pellets and nesting material. The study was approved by local ethics committee.
NOG mice were inoculated with 2.5 × 106 cells of the human melanoma cell line, FM82 (ESTDAB-27), subcutaneous (sc.) on the left flank. Tumor growth was measured with calipers twice a week, and represented as tumor surface (mm2). For studies of tumor homing, animals were treated with tail vein administration of 15 × 106 MAGE-A3 specific Mock or CXCR2 transduced T cells when tumor size reached approx. 30 mm2. ACT was combined with intra peritoneal (ip.) injection of 6 × 105 IU rh-IL2 at the day of ACT and twice daily for the subsequent two days. For study of tumor homing, blood was collected from the mandibular vein immediately prior to culling by cervical dislocation 7 days after ACT. Tumor, spleen, lungs and bone marrow were resected and kept in RPMI 1640 + 1% penicillin/streptomycin (Gibco) on ice until processing.
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3

Establishment of Colorectal Tumor PDX Models

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Tumor and matched normal specimens were obtained from patients undergoing liver metastasectomies or colon resections of primary disease at the University of Michigan University Hospital (Ann Arbor, MI). All patients provided informed written consent and samples were procured with approval of the University of Michigan Institutional Review Board (HUM00065489). Specimens were obtained within four hours of surgery and immediately transferred to DMEM/F12 media supplemented with 10% fetal bovine serum and 1% Penicillin-Streptomycin-Glutamine at 4°C. A portion of normal colon specimens were fixed in 10% neutral buffered formalin (NBF) and the remainder snap frozen in liquid nitrogen. Portions of tumor specimens were either fixed in 10% NBF, snap frozen in liquid nitrogen, or divided into fragments approximately 3 × 3 mm for subcutaneous implantation into female 6–7 week old CIEA NOG mice (NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac from Taconic) using an 11G Trocar needle. Tumor-implanted mice were monitored for tumor growth for up to four months following implantation. Xenografted tumors from the NOG mice were passaged into female 6–7 week old NCR nude mice (CrTac:NCr-Foxn1nu from Taconic) for model expansion. PDX models were maintained in nude mice for no more than four passages before fresh material from the freezer was used to regenerate the line.
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4

Establishing Patient-Derived Xenograft Models

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Surgical samples and blood were collected with informed consent under an Institutional Review Board-Approved Protocol. Following collection, the first 13 PDXs were established by implanting tumor fragments to the flank of nude mice as previously described (13 (link)). The latter models were established using a similar surgical procedure but involved placing the fragment on the bilateral 4th mammary fat pads of female nude mice (Department of Radiation Oncology-MD Anderson Cancer Center) or NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac (Taconic). Estrogen supplementation was provided to one of the specimens BCX.066 as it was 60% ER+ during presurgery clinical pathological analysis. All animal experiments were approved by Institutional Animal Care and Use Committee.
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5

Xenograft Transplantation of Human HSPCs

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Six- to 8-wk-old female CIEA NOG mice (NOD.Cg PrkdcscidIl2rgtm1Sug/JicTac) were purchased from Taconic Biosciences. One day after electroporation, 1 × 105 cells were administered by tail-vein injection after sublethal X-ray irradiation (75 cGy). Before transplantation, flow cytometry confirmed that >98% of HSPCs were CD34+. Mice were randomly assigned to each experimental group and evaluated in a blinded fashion.
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6

Xenograft Tumor Growth Measurement

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Mouse experiments were approved by the Landesamt für Gesundheit und Soziales Berlin (Reg 0010/19) and complied with the German Animal Welfare Act. B cell lines frozen 60 days after transduction were thawed and cultured for 2 weeks. 100 μl containing 5 million cells in 50% MatriGel (Corning, Cat. No. 356234) were injected subcutaneously into female, 2-month-old NOG mice (NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac; Taconic Biosciences). Three times per week the tumor volume was measured and calculated using the formula 0.5 a × b2, where a and b are the long and short diameters of the tumor, respectively. After reaching a tumor volume of ≥ 1.5 cm3 animals had to be sacrificed (humane endpoint).
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7

Xenograft Model of Leukemia

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All animal experiments were performed with the approval of Dana-Farber Cancer Institute's Institutional Animal Care and Use Committee. NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac (NOG) mice were obtained from Taconic Biosciences. Nonirradiated 8- to 12-week-old adult mice were transplanted with previously established PDXs (25 (link)) via tail-vein injection (250,000 cells/mouse). Engraftment of human cells (hCD45+) was analyzed and monitored longitudinally by weekly bleeding to quantify hCD45+ cells in the peripheral blood by flow cytometry with human CD45-PE and anti-mouse CD45-APC-Cy7 antibodies (BioLegend). Mice were monitored closely to detect disease onset, and treatment started when hCD45+ cells were detectable in the peripheral blood. Mice were randomly assigned to either normal or 0.1% VTP-50469 rodent special diet (25 (link)). Mice were bled weekly to monitor leukemia burden as described above and euthanized when showing clinical signs of disease (experimental endpoint). Leukemia cells from a subset of these animals were harvested after 7 days of treatment to perform RNA-seq.
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8

Xenograft Mouse Model Development

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Six-week-old immunodeficient male mice (BALB/c nude, n=58, CAnN.Cg-Foxn1nu/Crl, Charles River, Germany; NOG mice, n=7, NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac Taconic, Denmark; NOD.SCID mice, n=6, NOD/MrkBomTac-Prkdc SCID, Taconic, Denmark; or SHrN hairless NOD.SCID, n=13, NOD.Cg-PrkdcscidHrhr/NCrHsd, Envigo, France, Table 7) were maintained under controlled pathogen-free conditions (20–21 °C, 30–60% relative humidity, 12-hour light cycle, soya-free chow, and autoclaved drinking water).
Tissue specimens were aseptically cut into 1–2 mm3 pieces in a laminar hood using razor blades. Tumor pieces were implanted subcutaneously into the inter-scapular fat pads of immunodeficient mice. A total of 33 mice were supplemented with 20–25 mg Te propionate pellets, which were pressed in-house from Te-propionate powder (Sigma-Aldrich). Nineteen mice were supplemented with 12.5 mg 5α-DHT pellets (Innovative Research of America, FL, USA). Thirty-two mice received no hormone pellets.
At sacrifice, gross necropsy was performed, and the findings were documented. Tumors were removed and collected for histology or transplanted further to second or third generation of mice using the same protocol as described above.
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9

Xenograft Model of B-ALL in NOG Mice

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NOG mice (NOD.Cg-PrkdcscidIl2rgtm1Sug/JicTac) were purchased from Taconic. All animal experiments were approved by Verona University Ethical Committee (http://www.medicina.univr.it/fol/main?ent=bibliocr&id=85) and authorized by Ministerial Decree (16/2014-B). B-ALL animal experiments were in accordance with the Amsterdam Protocol on animal protection and welfare and conducted according to the guidelines of Federation of European Laboratory Animal Science Associations (FELASA).
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10

Murine Models for Xenograft Studies

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C57BL/6 and C57BL/-Tg (HLA-A2.1) 1 Enge/J (6–8 week-old male), and NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ mice (6–8 week-old male), known as NOD scid gamma (NSG), were purchased from Jackson Laboratory. Functionally equivalent NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac (6–8 week-old male), known as NOG, and C.B-Igh-1b/IcrTac-Prkdcscid (6–8 week-old male), known as CB17 SCID, were purchased from Taconic. All studies were conducted in accordance with IACUC approved protocols.
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