Blood samples from healthy individuals were purchased from Interstate Blood Bank (Memphis, TN), collected in tubes containing ethylenediaminetetraacetic acid (EDTA) (Becton Dickinson 366643) or CellSave® Preservative Tubes (Janssen Diagnostics). Blood samples (4 mL for Parsortix™ and 7.5 mL for CellSearch®) from 26 patients were analyzed in this study [metastatic breast (n = 10), nonmetastatic colon (n = 5), metastatic colon (n = 5), metastatic lung (n = 5) and nonmetastatic lung (n = 1)]. Cancer patients, treated at the University Medical Center Hamburg‐Eppendorf, were collected into CellSave® Preservative Tubes (Janssen Diagnostics) and processed by Parsortix™ or CellSearch® within 96 hours. The study was carried out in accordance with the World Medical Association Declaration of Helsinki and the guidelines for experimentation with humans by the Chambers of Physicians of the State of Hamburg (“Hamburger Ärztekammer”). The experimental protocol was approved (Approval No. PVN‐3779) by the Ethics Committee of the Chambers of Physicians of the State of Hamburg (“Hamburger Ärztekammer”). All participants gave written informed consent before the study began. Samples were prepared in the same manner described in the materials and methods and in accordance with the operating procedures for the CellSearch® and Parsortix systems.
Cellsave preservative tube
Manufactured by Johnson & Johnson
Sourced in United States
The CellSave Preservative Tubes are laboratory equipment designed for the collection and preservation of blood samples. The tubes contain a proprietary preservative formulation that helps maintain the integrity of the collected cells during storage and transportation.
Automatically generated - may contain errors
Lab products found in correlation
Vacutainer glass mononuclear cell preparation tubes, by BD (2 mentions)
Edta tube, by BD (2 mentions)
Cellsearch system, by Johnson & Johnson (2 mentions)
K2edta tubes, by BD (1 mentions)
Vacutainer, by BD (1 mentions)
Cellsearch cxc kit, by Johnson & Johnson (1 mentions)
Celltracker green, by Thermo Fisher Scientific (1 mentions)
Cellsearch ctc kit, by Johnson & Johnson (1 mentions)
Celltracks analyzer 2, by Johnson & Johnson (1 mentions)
19 protocols using cellsave preservative tube
1
Comparative Liquid Biopsy Evaluation
2
Metastatic Breast Cancer Biomarker Protocol
3
Breast Cancer Patient Blood Sampling
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In total, whole peripheral blood samples were drawn prospectively from 36 women who were actively undergoing treatment for previously confirmed stage III or stage IV breast cancer, at either the Fox Chase Cancer Center (FCCC) or University of Maryland, Baltimore (UMB) between 2011 and 2013. The study group characteristics can be found in Additional file 1 : Table S1. Anonymized peripheral blood samples were supplied through a collaboration agreement with FCCC and UMB, with written informed consent and according to the local Institutional Review Board (IRB) approval at Fox Chase Cancer Center or University of Maryland Baltimore. In addition, healthy female volunteers (n = 16; median age 52 years), donated blood samples with written informed consent and approval from Western Institutional Review Board. All anonymized blood samples were drawn into CellSave preservative tubes™ (approximately 9 mL; Janssen Diagnostics): 7.5 mL of blood was used to enumerate CTCs using CellSieve™ microfiltration at UMB, FCCC or Creatv Microtech. Results and patient identification from institutions were not shared or communicated until completion of study.
4
Plasma Isolation from Blood Samples
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After obtaining written informed consent, 9 × 10 mL of blood samples was collected within a single blood draw (see Fig. S1 ). Matched blood samples were collected in sterile 3 × 10 mL K2EDTA tubes (ETDA) (BD Vacutainer®, Becton Dickinson, Franklin Lanes, NJ, USA), 3 × 10 mL Cell‐Free DNA™ BCT (BCT) (Streck Inc., Omaha, NE, USA), and 3 × 10 mL CellSave Preservative tubes (Janssen Diagnostics, Raritan, NJ, USA) according to the manufacturer's instructions. The blood samples from one of each type of tube (EDTA, BCT, and CellSave) were processed for plasma isolation at three different time points: within 1 h, at 24 h and at 96 h after blood draw (see Fig. S1 ). Plasma was isolated using two sequential centrifugation steps: (a) 1711 g for 10 min at room temperature and (b) 12 000 g for 10 min at room temperature. Plasma was stored at −80 °C in 1 mL aliquots immediately after centrifugation until further processing.
5
CellSearch and PFC Blood Sampling
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PB samples have been drawn from central catheter, in order to decrease risks of endothelial cell detachment due to traumatic damage from venipuncture. Samples for CellSearch count were collected in specifically dedicated tubes (CellSave Preservative Tubes, Janssen Diagnostics LLC, Raritan, NJ, USA), that guarantee the reproducibility of results up to 96 hours from blood drawn; while samples for PFC count were collected in three EDTA (2 mg/ml) tubes (BD K2E EDTA, Becton Dickinson Biosciences - BD, San Jose, CA, USA). Leukocyte count, determined on each first drawn tube, was used for double platform calculation.
6
Spiking of Cell Lines into Blood
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Spiking was performed by adding 1 million MCF7 or SKBR3 cells into 7.5 mL blood in CellSave preservative tubes (Janssen Diagnostics, LLC, Raritan, NJ, USA). In all, 100,000 JEG-3 cells were spiked into 9 mL blood sample in Greiner Vacuette tubes (Greiner Bio-One GmbH, Germany). The blood tubes were immediately processed. The blood was sampled from healthy donors (Nordsjaellands Hospital, Hillerød, Denmark, or Ghent University, Belgium). A written informed consent in accordance with the Declaration of Helsinki was obtained from each participant before sample collection.
7
Metastatic Lung Cancer Blood Samples
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Peripheral blood samples were drawn by venipuncture into 10 mL CellSave Preservative Tubes (Janssen Diagnostics, Huntingdon Valley, PA, USA) from healthy donors and metastatic lung cancer patients treated at the University Medical Center Groningen. Patient demographics are provided in Table 2 . The experiments where done in accordance with relevant guidelines. All patients provided written informed consent and the study protocol was approved by the medical ethical committee of the University Medical Center Groningen (Groningen, The Netherlands). Healthy volunteers aged 20–55 gave written informed consent before donating blood.
8
Capturing Pancreatic Tumor Cells from Blood
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Five treatment-naive patients with biopsy-proven pancreatic ductal adenocarcinoma were enrolled for analysis after informed consent was obtained (UM IRB HUM00025339). Blood was collected and analyzed, as previously described48 (link). Briefly, peripheral venous blood was obtained through venipuncture and collected into Cell Save preservative tubes (Janssen Pharmaceuticals, Raritan, New Jersey). One milliliter was applied via microfluidic pump to a geometrically enhanced differential immunocapture (GEDI) chip functionalized previously with capture antibodies specific to epithelial cell adhesion molecule (EpCAM). After application of blood, the chip was washed with running buffer and antibodies to Cytokeratin and Synaptophysin conjugated to fluorophores and DAPI were applied to stain cells captured on the GEDI chip, as previously described. Stained cells were counted blindly as previously described. Scale bars indicate 100 microns.
9
Blood Sample Collection and Processing
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Healthy blood donors donated 20 mL of blood, collected either in 2 × 10 mL CellSave preservative tubes (Janssen Diagnostics, Raritan, NJ, USA) or in 1 × 10 mL EDTA tube (Becton, Dickinson and Company, Franklin Lakes, NJ, USA) and 1 × 10 mL CellSave preservative tube. Patients donated 3 × 10 mL of blood collected in CellSave preservative tubes. Blood samples were stored at room temperature until further processing. After blood draw, samples in EDTA tubes were processed within 24 h, whereas samples in CellSave tubes were processed within 96 h for plasma isolation as previously described (van Dessel et al., 2017 ).
10
Blood Collection for Cell Analysis
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7.5 mL of whole blood were collected into CellSave Preservative tubes (Janssen) containing EDTA and a cell fixative from patients and healthy normal volunteer donors, prior informed consent. Samples were maintained at room temperature and processed within 96 h. The methods were carried out in accordance with relevant guidelines. The study protocol was approved by the local ethical Committee of the Policlinico Umberto I, Sapienza University of Rome (Italy), protocol n. 668/09. Informed consent was obtained from all patients.
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