Large volume whole blood mononuclear cell enrichment using a COBE spectra

Cytotherapy ◽  
2013 ◽  
Vol 15 (4) ◽  
pp. S50
Author(s):  
N. Almezel ◽  
S. Pinkard ◽  
T. Leemhuis
2006 ◽  
Vol 13 (6) ◽  
pp. 704-707 ◽  
Author(s):  
Marcin Moniuszko ◽  
Krzysztof Kowal ◽  
Malgorzata Rusak ◽  
Miroslawa Pietruczuk ◽  
Milena Dabrowska ◽  
...  

ABSTRACT We investigated whether the choice of anticoagulant or the application of density gradient mononuclear cell isolation may account for conflicting published data regarding the levels of the scavenger receptors' expression in healthy individuals. We demonstrate that the detection of CD163, but not CD36, differs dramatically among the methods.


2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Ling Lin ◽  
Yong-Song Yue ◽  
Ni-Dan Wang ◽  
Lei-Yan Wei ◽  
Yang Han ◽  
...  

Abstract Background A more time saving, convenient, reproducible, and scalable method is needed to assess total HIV-1 DNA levels. Methods Frozen whole blood and peripheral blood mononuclear cell (PBMC) samples both 200 μl at the same point were used to detect total HIV-1 DNA. Automatic extraction of total HIV-1 DNA was used to ensure the consistency of sample extraction efficiency. The detection reagent was HIV-1 DNA quantitative detection kit and real-time quantitative PCR was utilized. Results Of the 44 included patients, 42 were male and 2 were female, with a median age of 33 years. Thirty-three cases were collected after receiving antiviral treatment, with a median duration of treatment of 3 months, and the other 11 cases were collected before antiviral treatment. The median viral load was 1.83 log10 copies/mL, the median CD4 and CD8 count were 94 and 680 cells/μL, and the median CD4/CD8 ratio was 0.18. The results of the two samples were 3.02 ± 0.39 log10 copies/106 PBMCs in PBMC samples and 3.05 ± 0.40 log10 copies/106 PBMCs in whole blood samples. The detection results of the two methods were highly correlated and consistent by using paired t test (P = 0.370), pearson correlation (r = 0.887, P < 0.0001) and intra-group correlation coefficient (ICC = 0.887, P < 0.0001) and bland-altman [4.55% points were outside the 95% limits of agreement (− 0.340 ~ 0.390)]. Conclusions The results of the whole blood sample test for total HIV-1 DNA are consistent with those of PBMC samples. In a clinical setting it is recommended to use whole blood samples directly for the evaluation of the HIV reservoir.


Blood ◽  
1998 ◽  
Vol 91 (11) ◽  
pp. 4216-4223 ◽  
Author(s):  
Nigel S. Key ◽  
Arne Slungaard ◽  
Luke Dandelet ◽  
Stephen C. Nelson ◽  
Christopher Moertel ◽  
...  

Abstract We developed a simple assay for the measurement of tissue factor procoagulant activity (TF PCA) in whole blood samples that avoids the need for mononuclear cell isolation. This method combines convenience of sample collection and processing with a high degree of sensitivity and specificity for TF. Using this method, we have determined that TF PCA is detectable in whole blood samples from normal individuals, which is itself a novel observation. Essentially all PCA could be shown to be localized in the mononuclear cell fraction of blood. Compared with controls, whole blood TF levels were significantly (P &lt; .000001) elevated in patients with sickle cell disease (SCD), regardless of the subtype of hemoglobinopathy (SS or SC disease). No significant difference in TF PCA was observed between patients in pain crisis compared with those in steady-state disease. Because TF functions as cofactor in the proteolytic conversion of FVII to FVIIa in vitro, it was expected that an increase in circulating TF PCA would lead to an increased in vivo generation of FVIIa. On the contrary, FVIIa levels were actually decreased in the plasma of patients with SCD. Plasma TF pathway inhibitor (TFPI) antigen levels were normal in SCD patients, suggesting that accelerated clearance of FVIIa by the TFPI pathway was not responsible for the reduced FVIIa levels. We propose that elevated levels of circulating TF PCA may play an important role in triggering the activation of coagulation known to occur in patients with SCD. Because TF is the principal cellular ligand for FVIIa, it is possible that increased binding to TF accounts for the diminished plasma FVIIa levels.


1986 ◽  
Vol 250 (3) ◽  
pp. E331-E337
Author(s):  
H. R. Berthoud ◽  
W. B. Laughton ◽  
T. L. Powley

A new protocol that makes it feasible to withdraw large volumes of whole blood from an individual rat within 1 h or less is described. This method involves the use of indwelling catheters for withdrawal of blood from the inferior vena cava with concurrent isovolemic replacement of whole blood into the superior vena cava. Simultaneity of the transfusion and withdrawal, strict equality of volumes, and a smooth exchange of blood are assured by the use of separate channels of the same multiple-channel peristaltic pump for withdrawal and replacement. Validation experiments using both anesthetized and unanesthetized rats indicate that several responses remain essentially undisturbed during large volume blood sampling; these parameters include blood pressure, heart rate, hematocrit, plasma hormones including insulin and glucagon, plasma glucose levels, and feeding behavior. Considerations of technical and physiological limitations of the protocol, including choice of catheters and catheter placement, pump, sampling parameters, and obtaining donor blood, are discussed.


Sign in / Sign up

Export Citation Format

Share Document