Peripheral blood was collected by venous puncture into Vacutainer Plus Plastic K3EDTA Tubes or Vacutainer CPT Mononuclear Cell Preparation Tubes with Sodium Heparin (BD Biosciences). Complete blood counts were monitored at all time points from the Vacutainer Plus Plastic K3EDTA Tubes. Plasma was isolated either from Vacutainer Plus Plastic K3EDTA Tubes by centrifugation for 10 min at 480 × g or from Vacutainer CPT Mononuclear Cell Preparation Tubes with Sodium Heparin after centrifugation for 40 min at 1900 × g, and the samples were stored at −80 °C. Peripheral blood mononuclear cells (PBMCs) were isolated from Vacutainer CPT Mononuclear Cell Preparation Tubes with Sodium Heparin according to the manufacturer’s instructions (BD Biosciences), and red blood cells were lysed in ammonium-chloride-potassium (ACK) buffer (0.15 M NH4Cl, 10 mM KHCO3, 0.1 mM EDTA, pH 7.4).
Vacutainer cpt mononuclear cell preparation tubes with sodium heparin
Manufactured by BD
Sourced in United States
The BD Vacutainer CPT Mononuclear Cell Preparation Tubes with Sodium Heparin are designed for the separation and isolation of mononuclear cells from whole blood samples. The tubes contain a gel and a density gradient medium to facilitate the separation of mononuclear cells during centrifugation.
Automatically generated - may contain errors
Lab products found in correlation
Vacutainer pet edta tubes, by BD (1 mentions)
Facs lsrfortessa, by BD (1 mentions)
Zombie yellow fixable viability kit, by BioLegend (1 mentions)
Anti cd56 apc, by Beckman Coulter (1 mentions)
Anti hladr v500, by BD (1 mentions)
Anti cd123 fitc, by BioLegend (1 mentions)
Anti cd19 apc h7, by BD (1 mentions)
Anti cd56 bv421, by BioLegend (1 mentions)
Anti cd3 percp cy5, by BioLegend (1 mentions)
Anti cd14 ecd, by Beckman Coulter (1 mentions)
Matrigel coated plate, by Corning (1 mentions)
Dneasy blood and tissue kit, by Qiagen (1 mentions)
Mtesr1 medium, by STEMCELL (1 mentions)
Bambanker, by Nippon Genetics (1 mentions)
6 protocols using vacutainer cpt mononuclear cell preparation tubes with sodium heparin
1
Peripheral Blood Collection and Processing
2
Monocyte Isolation and ROS Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human subjects’ study was sponsored by the University Hospital of Toulouse for regulated and ethical submission (Clinical trials.gov NCT01990716). Human blood was obtained from IBD patients and collected in BD Vacutainer® CPT™ Mononuclear Cell Preparation Tubes with Sodium Heparin (BD Biosciences). Mononuclear cells were isolated following the manufacturer instructions and monocytes were isolated by adherence to plastic for 2 h in SFM (Gibco) at 37 °C, 5% CO2. Monocytes were then plated for reactive oxygen species (ROS) release assessment, and gene and protein expression after addition of Naticol®Gut (100 µg/ml) for 6 h.
3
Isolation and Cryopreservation of PBMCs and Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs from healthy donors and participants were isolated from fresh blood samples using BD Vacutainer® CPT™ Mononuclear Cell Preparation Tubes (with Sodium Heparin, BD Cat# 362780) in a density gradient centrifugation at the same day of blood collection. Afterwards, PBMCs were cryopreserved in freezing medium (90% of fetal bovine serum (FBS) + 10% dimethyl sulfoxide) in liquid nitrogen until further use. Plasma samples were obtained using BD Vacutainer™ PET EDTA Tubes centrifugation, aliquoted and cryopreserved at −80 °C until further use.
4
Isolation and Characterization of Plasmacytoid Dendritic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh PBMCs were isolated from whole blood by density-gradient centrifugation using the BD Vacutainer CPT Mononuclear Cell Preparation Tubes with Sodium Heparin (BD Biosciences, Franklin Lakes, NJ, USA). The cells were first stained using the Zombie Yellow™ Fixable Viability Kit (BioLegend, San Diego, CA, USA) and then with combinations of the following monoclonal antibodies against human cell-surface antigens for 30 min on ice: anti-CD11c-Alexa700, anti-HLADR-V500, anti-CD19-APC-H7 (all from BD Biosciences), anti-CD14-ECD, anti-CD56-APC, (both from Beckman Coulter, Brea, CA, USA), anti-CD123-FITC, anti-CD3- PerCPCy5.5, anti-CD56-BV421 (all from BioLegend), and anti-CD19-PE (TONBO Biosciences, San Diego, CA, USA). pDCs were identified as CD3-CD19-CD14-CD56-HLADR+CD11c-CD123+(Additional file 2 : Figure S1). Data were acquired on a FACS LSR Fortessa (BD Biosciences) and the percentages of each cell population and mean fluorescence intensity were analyzed using FlowJo software (TreeStar Inc., Ashland, OR, USA).
5
Efficient hiPSC Generation from PBMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples were collected from all participants and used for both DNA isolation (DNeasy Blood and Tissue Kit, QIAGEN) and peripheral blood mononuclear cells (PBMCs) extraction (BD Vacutainer CPT Mononuclear Cell Preparation Tubes with Sodium Heparin, BD Biosciences) according to manufacturer’s instructions. PBMCs were cultured for 9 to 12 days in an erythroblast enrichment medium [42 ] to expand the erythroblast population and 2.5 × 105 cells were transduced using recombinant Sendai viral vectors (Cytotune-iPSC 2.0TM, Thermofisher scientific), expressing the four reprogramming factors Oct4, Sox2, Kfl4, and c-Myc, according to manufacturer’s instructions. After 3 days, transduced cells were plated on irradiated mouse embryonic fibroblast (MEFs; Fisher A34961) and grown for 2 to 3 weeks in hESC medium until the emergence of individual colonies. Live hiPSCs were labeled by Tra-1-60 immunostaining (R&D systems) and positive clones were manually picked and grown on MEFs using hiPSC hiPSC medium (DMEM/F12, 20% knockout serum, 1X MEM-NEAA, 1X L-glutamine, 1X pen/strep, 0.1 mM 2-mercaptoethanol, 20 ng/mL b-FGF). After reaching passage 10, hiPSC colonies were transitioned to feeder-free conditions using Matrigel-coated plates (Corning) and mTeSR1 medium (Stem Cell Technology) and up to three clones per individual were validated, expanded and cryopreserved.
6
Isolation and Cryopreservation of PBMCs from Ulcerative Colitis Patients
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Six patients with UC (Table 1 ) participated in this study. The clinical activity index of patients was evaluated using the Lichtiger index, and the endoscopic severity was determined using the Ulcerative Colitis Endoscopic Index of Severity. Peripheral blood was drawn after obtaining informed consent in accordance with the local ethical committee guidelines of Juntendo University. PBMCs were isolated from whole blood by density-gradient centrifugation using BD Vacutainer CPT Mononuclear Cell Preparation Tubes with Sodium Heparin (BD Biosciences). PBMCs were cryopreserved in cryopreservation medium (Bambanker; Nippon Genetics, Tokyo, Japan) until used.
Characteristics of the Patients Included in the Study
| Characteristic | Patients with UC (n = 6) |
|---|---|
| Mean age ± SD, y | 47.7 ± 13.3 |
| Sex (male/female) | 4/2 |
| Mean duration of disease ± SD, y | 12.0 ± 11.6 |
| Disease location, n | |
| Proctitis | 2 |
| Left-sided colitis | 1 |
| Extensive colitis | 3 |
| Ongoing treatments, n | |
| 5-ASA | 5 |
| Azathioprine, 6-MP | 2 |
| Anti-TNF therapy | 2 |
| Anti-α4β7 integrin therapy | 1 |
| PSL | 2 |
| Median Lichtiger index (range) | 1 (0-1) |
| Median UCEIS (range) | 1.5 (0-3) |
| Mean CRP ± SD, mg/dL | 0.17 ± 0.21 |
5-ASA, 5-Aminosalicylic acid; CRP, C-reactive protein; 6-MP, mercaptopurine; PSL, prednisolone; SD, standard deviation; TNF, tumor necrosis factor; UC, ulcerative colitis; UCEIS, Ulcerative Colitis Endoscopic Index of Severity.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!