Subcutaneous tumours were generated in 6- to 7-week-old male Athymic Nude Foxn1nu/nu mice (ENVIGO) by injecting 2 × 106 MIA PaCa-2 or PANC-1 cells embedded in Matrigel 1:1 (BD Biosciences) into each flank. Tumours were measured at least twice weekly, and the tumoural volumes calculated using the formula, V = D × d2 × π ÷ 6 V. Mice were randomly assigned to either group for treatment. Viruses (4 × 1010 vp/animal) were administered intravenously in physiological saline solution once tumours achieved a median volume of 100 mm3. The experiment remained blinded until the conclusion of the study. Animals were euthanized 45–48 days after virus administration, and tumours were collected and flash-freezed in liquid nitrogen. All animal procedures met the guidelines of European Community Directive 86/609/EEC and were approved by the ethical committee (CEEA-University of Barcelona) and by the local authorities of the Generalitat de Catalunya.
Matrigel 1 1
Manufactured by BD
Sourced in United States
Matrigel 1:1 is a reconstituted basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, a tumor rich in extracellular matrix proteins. It is provided as a 1:1 dilution in Dulbecco's Modified Eagle Medium (DMEM).
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using matrigel 1 1
1
Subcutaneous Tumor Induction and Virotherapy
2
Xenograft Tumor Experiments in NSG Mice
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Xenograft tumor experiments were performed in 5-week-old NOD SCID-gamma (NSG) mice. The animal study was approved by the Institutional Animal Care and Use Committees of Hanyang University. All mice were housed in a temperature-controlled room under standard conditions (12 h light/dark cycle and 55% relative humidity) with access to food and water ad libitum. SH-SY5Y cells transfected with mock control, USP3-KO, and USP3-KO clones reconstituted with USP3 or REST cells were prepared in DMEM: Matrigel (1:1) (BD Biosciences) and subcutaneously injected into the right flank of each mouse. The tumors were harvested at the end of the experiment and images were taken. Tumor growth was recorded by measuring two perpendicular diameters (short axis and long axis), and tumor volume was calculated using the formula V = D × d2 × 0.5, where D and d are the long and short axes of the tumor, respectively. For all animal studies, the animals were chosen randomly.
3
Xenograft Model for Pancreatic Cancer
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RWP-1, MIA PaCa-2 and PANC-1 cells (2.5 × 106), embedded in Matrigel 1:1 (BD Biosciences, San Jose, CA, USA), were subcutaneously injected into each flank of male, 7- to 8-week-old, athymic nu/nu mice (Harlan, Sant Feliu de Codines, Spain). Tumours were measured at least three times weekly, and their volumes were calculated using the formula V=larger diameter × (smaller diameter)2 × pi÷6. Mice were randomly assigned to either group for treatment. Virus was administered once tumours achieved a median volume of 100 mm3. The experimenter was blinded until the conclusion of the study.
4
Subcutaneous Xenograft Mouse Model for PDAC
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RWP-1 cells (2.5×106), embedded in Matrigel 1:1 (BD Biosciences, San Jose, CA), were injected subcutaneously into each flank of male, 6-8-weeks-old, athymic nu/nu mice (Harlan, Sant Feliu de Codines, Spain). PDAC fragments of 10-15 mg from CP15 and CP13 patient-derived xenografts (PDX), patient specimens maintained as xenografts in the mouse pancreas [28 (link), 29 (link)] were implanted in the subcutaneous tissue of 7-weeks-old, athymic nu/nu mice.
Viral treatment was administered at day 9-(RWP-1), 7-(CP13) and 15-(CP15) post implantation when tumors achieved a median volume of 170, 100 and 80 mm3 respectively. Tumors were measured at least three times a week and their volume was calculated by the formula V = larger diameter × (smaller diameter)2 × pi ÷ 6. At the end of the experiment tumor weight was calculated for CP13 and CP15 PDX. All animal procedures met the guidelines of European Community Directive 86/609/EEC, and approved by the local ethical committee.
Viral treatment was administered at day 9-(RWP-1), 7-(CP13) and 15-(CP15) post implantation when tumors achieved a median volume of 170, 100 and 80 mm3 respectively. Tumors were measured at least three times a week and their volume was calculated by the formula V = larger diameter × (smaller diameter)2 × pi ÷ 6. At the end of the experiment tumor weight was calculated for CP13 and CP15 PDX. All animal procedures met the guidelines of European Community Directive 86/609/EEC, and approved by the local ethical committee.
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