scholarly journals High-throughput, low-loss, low-cost, and label-free cell separation using electrophysiology-activated cell enrichment

2017 ◽  
Vol 114 (18) ◽  
pp. 4591-4596 ◽  
Author(s):  
Shabnam A. Faraghat ◽  
Kai F. Hoettges ◽  
Max K. Steinbach ◽  
Daan R. van der Veen ◽  
William J. Brackenbury ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Cancer Cells ◽  
Blood Cells ◽  
Cell Separation ◽  
Low Cost ◽  
Operating Cost ◽  
Gradient Methods ◽  
Cell Loss ◽  
Cell Recovery ◽  
Processing Rate

Currently, cell separation occurs almost exclusively by density gradient methods and by fluorescence- and magnetic-activated cell sorting (FACS/MACS). These variously suffer from lack of specificity, high cell loss, use of labels, and high capital/operating cost. We present a dielectrophoresis (DEP)-based cell-separation method, using 3D electrodes on a low-cost disposable chip; one cell type is allowed to pass through the chip whereas the other is retained and subsequently recovered. The method advances usability and throughput of DEP separation by orders of magnitude in throughput, efficiency, purity, recovery (cells arriving in the correct output fraction), cell losses (those which are unaccounted for at the end of the separation), and cost. The system was evaluated using three example separations: live and dead yeast; human cancer cells/red blood cells; and rodent fibroblasts/red blood cells. A single-pass protocol can enrich cells with cell recovery of up to 91.3% at over 300,000 cells per second with >3% cell loss. A two-pass protocol can process 300,000,000 cells in under 30 min, with cell recovery of up to 96.4% and cell losses below 5%, an effective processing rate >160,000 cells per second. A three-step protocol is shown to be effective for removal of 99.1% of RBCs spiked with 1% cancer cells while maintaining a processing rate of ∼170,000 cells per second. Furthermore, the self-contained and low-cost nature of the separator device means that it has potential application in low-contamination applications such as cell therapies, where good manufacturing practice compatibility is of paramount importance.

scholarly journals Effects of gadolinium-based magnetic resonance imaging contrast media on red blood cells and K562 cancer cells

2020 ◽  
Vol 62 ◽  
pp. 126640
Author(s):  
Benjamaporn Supawat ◽  
Phattharawadi Moungthong ◽  
Chananchida Chanloi ◽  
Natchaporn Jindachai ◽  
Singkome Tima ◽  
...  
Keyword(s):  
Magnetic Resonance Imaging ◽  
Magnetic Resonance ◽  
Contrast Media ◽  
Red Blood Cells ◽  
Cancer Cells ◽  
Blood Cells ◽  
Resonance Imaging ◽  
Magnetic Resonance Imaging Contrast

Combined Platelet and Erythrocyte Salvage: Evaluation of a New Filtration-based Autotransfusion Device

2021 ◽  
Author(s):  
Alexandre Mansour ◽  
Benoit Decouture ◽  
Mikaël Roussel ◽  
Charles Lefevre ◽  
Lucie Skreko ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Whole Blood ◽  
Blood Cells ◽  
Plasma Proteins ◽  
Complete Blood Count ◽  
Cell Recovery ◽  
Healthy Human ◽  
Platelet Recovery ◽  
Human Whole Blood ◽  
And Function

Background The SAME device (i-SEP, France) is an innovative filtration-based autotransfusion device able to salvage and wash both red blood cells and platelets. This study evaluated the device performances using human whole blood with the hypothesis that the device will be able to salvage platelets while achieving a erythrocyte yield of 80% and removal ratios of 90% for heparin and 80% for major plasma proteins without inducing signification activation of salvaged cells. Methods Thirty healthy human whole blood units (median volume, 478 ml) were diluted, heparinized, and processed by the device in two consecutive treatment cycles. Samples from the collection reservoir and the concentrated blood were analyzed. Complete blood count was performed to measure blood cell recovery rates. Flow cytometry evaluated the activation state and function of platelets and leukocytes. Heparin and plasma proteins were measured to assess washing performance. Results The global erythrocyte yield was 88.1% (84.1 to 91.1%; median [25th to 75th]) with posttreatment hematocrits of 48.9% (44.8 to 51.4%) and 51.4% (48.4 to 53.2%) for the first and second cycles, respectively. Ektacytometry did not show evidence of erythrocyte alteration. Platelet recovery was 36.8% (26.3 to 43.4%), with posttreatment counts of 88 × 109/l (73 to 101 × 109/l) and 115 × 109/l (95 to 135 × 109/l) for the first and second cycles, respectively. Recovered platelets showed a low basal P-selectin expression at 10.8% (8.1 to 15.2%) and a strong response to thrombin-activating peptide. Leukocyte yield was 93.0% (90.1 to 95.7%) with no activation or cell death. Global removal ratios were 98.3% (97.8 to 98.9%), 98.2% (96.9 to 98.8%), and 88.3% (86.6 to 90.7%) for heparin, albumin, and fibrinogen, respectively. The processing times were 4.4 min (4.2 to 4.6 min) and 4.4 min (4.2 to 4.7 min) for the first and second cycles, respectively. Conclusions This study demonstrated the performance of the SAME device. Platelets and red blood cells were salvaged without significant impact on cell integrity and function. In the meantime, leukocytes were not activated, and the washing quality of the device prevented reinfusion of high concentrations of heparin and plasma proteins. Editor’s Perspective What We Already Know about This Topic What This Article Tells Us That Is New

scholarly journals Dual secured nano-melittin for the safe and effective eradication of cancer cells

2015 ◽  
Vol 3 (1) ◽  
pp. 25-29 ◽  
Author(s):  
Bei Cheng ◽  
Bindu Thapa ◽  
Remant K. C. ◽  
Peisheng Xu
Keyword(s):  
Red Blood Cells ◽  
Cancer Cells ◽  
Blood Cells ◽  
Wide Spectrum ◽  
Lytic Activity ◽  
Anticancer Efficacy

A dual secured nano-melittin system fully retains the wide-spectrum anticancer efficacy of melittin while quenching its lytic activity for the red blood cells.

scholarly journals Different responses of normal cells (red blood cells) and cancer cells (K562 and K562/Dox cells) to low-dose 137Cs gamma‑rays

2021 ◽  
Vol 14 (4) ◽  
Author(s):  
Benjamaporn Supawat ◽  
Panumas Homnuan ◽  
Natthawan Kanthawong ◽  
Niyada Semrasa ◽  
Singkome Tima ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Cancer Cells ◽  
Gamma Rays ◽  
Blood Cells ◽  
Low Dose ◽  
Normal Cells

Hematopoietic Stem Cell Transplantation (HSCT) for Benign Indications Using Umbilical Cord Blood Units (UCB) That Were Depleted of Plasma, but Not of Red Blood Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5459-5459
Author(s):  
Robert Chow ◽  
Tang-Her Jaing ◽  
David Gjertson ◽  
Joseph Rosenthal ◽  
Auayporn Nademanee ◽  
...  
Keyword(s):  
Stem Cell ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Acute Gvhd ◽  
Chronic Gvhd ◽  
Disease Free Survival ◽  
Critical Factor ◽  
Cell Loss ◽  
Hematopoietic Stem ◽  
Benign Disorders

Abstract Background: UCB is an attractive unrelated source for HSCT of benign indications such as hemoglobinapathies, genetic diseases and metabolic disorders, because of less stringent HLA matching requirements, lower incidence and severity of GvHD, and ready availability of a physical inventory; however, unlike BMT or PSCT, cell dosage is a critical factor in success of UCB HSCT. The red cell depletion (RCD) techniques that are widely used by UCB banks incur significant nucleated cell loss after processing. One method of minimizing cell loss during processing is to deplete plasma, but not the red blood cells (PD). A large racially diverse PD UCB inventory of 18,000 units is now available on stem cell registries. We now report our observations of using unrelated PD UCB HSCT for benign indications. Hypothesis: Usage of unrelated PD UCB for HSCT of benign indications will result in acceptable clinical outcome for myeloid (ANC 500) and platelet engraftment, overall survival (OS), and transplant related mortality (TRM). Methods: A retrospective analysis was performed on 28 patients with benign disorders who were transplanted with PD UCB in 15 centers and 7 countries, with 13 thalassemias, 5 SAA, 5 WAS, 2 SCID, 1 sickle cell disease, 1 osteopetrosis and 1 metabolic disorder. Results: The median age of patients was 3.7 years old (range 03–27 years old); median weight 16.0 kg (range 4.5–43 kg); male 57%. Transplant characteristics indicated a median # HLA ABDR matches of 4.0 (5 6/6; 7 5/6; 12 4/6; 3 3/6; 1NA); median pre-freeze TNC dose 7.7 x 107/kg; median post-thaw TNC dose as reported by TC 7.7 x 107/kg; median pre-freeze CD34 dose 3.1 x 105/kg; transplants performed outside of U.S. 68%; double unit transplant 13%; non-myeloablative 6%. The median time to engraftment for ANC 500 (n=21), platelet 20K (n=20), and 50K (n=18) are 14.5 days (range 11–41 days), 47.0 days (range 13–82 days), and 55.0 days (range 21–96 days) respectively. The unadjusted cumulative incidence (C.I.) of ANC500 and platelet 20K and 50K engraftment are 89±7%, 89±7%, and 87±8% respectively. The incidence of reported grade II–IV acute GVHD was 33%, and none had grade III–IV acute GVHD. 50% developed limited chronic GVHD (7/14), and so far only one patient was reported to have extensive chronic GVHD. With a median follow-up of 356 days (range 93–1,100 days), the Kaplan-Meier estimates of 1-year TRM, OS and disease-free survival were 11±6%, 89±6% and 89±6% respectively. Conclusion: These results demonstrate that HSCT using unrelated PD UCB can be performed safely and effectively in patients with benign disorders. Further study of HSCT using unrelated PD UCB in patients with non-malignant hematological disorders is warranted.

scholarly journals Blood Cell Separation in the Dog by Continuous Flow Centrifugation

Blood ◽  
1968 ◽  
Vol 31 (5) ◽  
pp. 653-672 ◽  
Author(s):  
DEAN BUCKNER ◽  
ROBERT EISEL ◽  
SEYMOUR PERRY
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Continuous Flow ◽  
Cell Separation ◽  
Mononuclear Cells ◽  
Buffy Coat ◽  
Donor Blood ◽  
The Mean ◽  
Blood Volumes

Abstract A closed continuous flow centrifuge was used to separate and collect large quantities of buffy coat cells from the dog. One hundred fifty-five separate centrifugations of 2-12 hours duration were performed. Up to 61.0 liters of blood, representing 2.2-52 donor blood volumes, were processed. Buffy coat cells with a preponderance of granulocytes or lymphocytes, were collected while plasma and red blood cells were returned to the donor without change in flow or gravitational field. The mean total number of leukocytes, granulocytes, mononuclear cells and platelets removed was 24.0, 17.0, 7.0 and 197.0 x 10,9 respectively, which was 34.0, 28.0, 66.0 and 36.0 percent, respectively, of each cell population entering the centrifuge. The effect of centrifugation on blood components was evaluated. Granulocytes from buffy coat collections exhibited normal phagocytic ability in vitro. The transfusion of large quantities of granulocytes (15.0 x 109) into leukopenic dogs produced an increment in peripheral granulocyte count (2.7 x 103 per mm.3). Granulocytes labeled with 3H-DFP were also infused with a recovery of 38.6 percent at one hour and a T-½ of 4-6 hours. Machine-separated lymphocytes responded normally to phytohemagglutinin. Prolonged passage of blood through the pumps, tubing and bowl without centrifugation resulted in a decrease in circulating platelet levels (21 percent). A greater decrease in platelets occurred with centrifugation at high g. forces (49 percent). Hemolysis of red blood cells was not a serious problem.

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