Sample collection
tumor
STERILE, TUMOR, METASTASIS, METASTASES
The enrolled patients underwent a biopsy of the metastasis (liver in > 90% of cases) based on the safety of the procedure and the accessibility of the lesion. All available imaging was evaluated to select active metabolic metastases with size progression, activity on fluorodeoxyglucose positron emission tomography, or diffusion restriction on magnetic resonance imaging, but only a computerized tomography scan (CT) was mandatory. Prior to the biopsies, all patients had a blood sample drawn to evaluate hemoglobin (Hgb) and coagulation parameters. The biopsy levels were set at Hgb concentration > 5 mmol/L, coagulation factor normal test (coagulation factor II, VII, X) > 0.4, activated partial thromboplastin time < 40 s. and thrombocytes > 40 × 10Image-guided biopsies were performed by an experienced radiologist using a SuperCore™ Semi-Automatic Biopsy Instrument (Argon Medical Devices, Inc.,1445 Flat Creek Road Athens, Texas 75,751 USA). An Aplio i800 ultrasound unit (Canon Medical Systems Corporation, 1385 Shimoishigami, Otawara-shi, Tochigi 324–8550, Japan) with a needle guide system (Verza™ Guidance System, CIVCO Medical Instruments Co., Inc. 102 First Street South, Kalona, IA, USA) was used for liver biopsy. CT-guided biopsies were performed with a CT-guided “beam-through” technique using a 64-slice CT scanner (Phillips Brilliance 64, Eindhoven, Netherlands). Re-biopsy was allowed if the patient was in good clinical performance. One to three (preferably) 18G to 16G (preferably) biopsies were sampled in a collection tube containing sterile phosphate buffered saline, 500 U/mL penicillin, 500 µg/mL streptomycin, and 5 mg/mL amphotericin. In parallel, a biopsy was formalin-fixed and paraffin-embedded, and a 4-micron section stained with Hematoxylin-Eosin was reviewed to confirm the presence of viable tumor cells.