{"content":[{"text":"NCI Patient-Derived Models Repository (PDMR) Material Request Procedures","bbox":{"left":0.106275,"top":0.046726,"width":0.6225269999999999,"height":0.016778999999999995}},{"text":"Effective Date:","bbox":{"left":0.106275,"top":0.072407,"width":0.11927,"height":0.016779000000000002}},{"text":"12/9/2020","bbox":{"left":0.253725,"top":0.072407,"width":0.07960800000000001,"height":0.016779000000000002}},{"text":"Page 2 of 5","bbox":{"left":0.711471,"top":0.072407,"width":0.0888230000000001,"height":0.016779000000000002}},{"text":"3.","bbox":{"left":0.147059,"top":0.105816,"width":0.014705999999999997,"height":0.016779000000000002}},{"text":"If requesting cryopreserved fragments for PDX generation, provide a copy of your Animal \n Care and Use Committee (ACUC) protocol indicating your laboratory, or designate, uses \n NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice. Alternatively, you may provide a Bill of \n Sale showing NSG mice have been purchased. All PDMR PDXs must be initially implanted \n into NSG mice for propagation. \n Requests will be processed after receipt of all required documentation; partial submissions \n will be returned. The requests will not be reviewed until all paperwork has been received. \n Please send the necessary paperwork to: NCI_PDM_Repository@mail.nih.gov","bbox":{"left":0.176471,"top":0.105816,"width":0.728227,"height":0.146325}},{"text":"4.","bbox":{"left":0.147059,"top":0.200513,"width":0.014705999999999997,"height":0.01677899999999999}},{"text":"Specimen types that may be requested: \n A single vial of the following material will be shipped upon arrival: \n • Vial of Cryopreserved PDX fragments: Single vial from a PDX tumor no higher than the \n listed maximum passage for the distribution lot; sufficient tissue to implant into 2-5 NSG \n mice. Material cannot be requested from a specific PDX sample in a lineage. \n • Vial of Flash-frozen DNA: Single vial from a PDX tumor no higher than the listed \n maximum passage for the distribution lot. Approximately 2-3 µg DNA in at least 10 µL \n prepared using Qiagen’s DNA/RNA AllPrep Mini Kit (cat#: 80204) \n • Vial of Flash-frozen RNA: Single vial from a PDX tumor no higher than the listed \n maximum passage for the distribution lot. Approximately 2-3 µg RNA in at least 10 µL \n prepared using Qiagen’s DNA/RNA AllPrep Mini Kit (cat#: 80204). RNA quality is \n periodically assessed using an Agilent 2100 Bioanalyzer and vials are maintained if the RIN \n is >5. For those interested in doing RNASeq, we would recommend requesting a flash- \n frozen fragment and performing an independent extraction. \n • Vial of Fresh-frozen PDX fragment: For protein extraction, single vial with a 30-60 mg \n fresh-frozen PDX tumor fragment no higher than the listed maximum passage for the \n distribution lot \n • Vial of Cryopreserved Patient/PDX-Derived Heterogenous Tumor Culture (PDC) cells: \n Vial contains a minimum of 7.5 x 105 cells. Cells are non-clonal, not transformed, and have \n been grown out to 20 passages maintained on Matrigel-coated surface in the recommended \n defined Media + Y compound without out-growth of fibroblasts or loss of fidelity. \n • Vial of Cryopreserved Patient-derived Cancer Associated Fibroblast Culture (CAF): \n Vial contains a minimum of 7.5 x 105 cells. Cells are non-clonal, not transformed, and have \n been grown for a minimum of 3 passages when maintained on Matrigel-coated surface in the \n recommended defined Media + Y compound. \n • Vial of Cryopreserved Patient/PDX-Derived Organoid (PDOrg) cells: Vial contains a \n minimum of 1-5 x 105 cells. Cells are non-clonal, non-transformed, and have been grown out \n to 10 passages maintained in the recommended defined Media + Y compound + Basement \n Membrane matrix according to the PDMR SOPs.","bbox":{"left":0.088235,"top":0.285362,"width":0.823291,"height":0.572839}}],"images_bbox":[{"left":0.088235,"top":0.918182,"width":0.390278,"height":0.03628},{"left":0.695098,"top":0.909091,"width":0.209314,"height":0.045453}],"images_bbox_not_text_overlap":[{"left":0.088235,"top":0.918182,"width":0.390278,"height":0.03628},{"left":0.695098,"top":0.909091,"width":0.209314,"height":0.045453}]}