Establishment of Patient-Derived Pancreatic Tumor Xenografts

Our use of the SCID mouse-patient tumor xenograft model has been previously described ([31 (link),33 (link),41 (link),32 (link),34 (link),30 (link)]). For these studies, surgical specimens of patients' pancreatic tumors were received shortly after resection through the Tissue Procurement Facility (TP) of RPCI and cut into 2 mm × 2 mm pieces in tissue culture medium (RPMI 1640) under sterile conditions. SCID mice were then anesthetized by intraperitoneal injection of 0.4–0.5 ml Avertin (2.5 g 2,2,2-tribromoethanol dissolved in 5 ml 2-methyl-butanol/200 ml ddH₂O) and individual tumor pieces were implanted subcutaneously in the abdominal wall of three mice (1^st passage) and monitored for growth. The mice used in all experiments were 7–8 weeks old CB17 SCID mice with an average weight of 18–20 g. They were kept in sterile cages (4–5 mice per cage) and fed with autoclaved chow and water. Mice were maintained in air-conditioned and light controlled rooms (12 hr cycles). All procedures, injections and tumor measurements were carried out under a laminar flow hood using aseptic precautions. Tumor specimens that grew to a size of 1 cm (8–12 weeks) were retrieved and subsequently passaged into recipient mice (2^nd passage) and were considered to have successfully engrafted when these tumors grew. Pathological diagnosis of patient specimens and evaluation of engrafted/ passaged tumors was performed in collaboration with a member of the Pathology Department at RPCI.