Clinical-scale elutriation as a means of enriching antigen-presenting cells and manipulating alloreactivity

Micklethwaite, Kenneth; Garvin, Frances; Kariotis, Melina; Yee, Leng L.; Hansen, Anna M.; Antonenas, Vicki; Sartor, Mary; Turtle, Cameron J.; Gottlieb, David

doi:10.1080/14653240802702160

Cited by 5 publications

(2 citation statements)

References 30 publications

(19 reference statements)

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“…However, almost all neutrophils were also found in fraction 5 with monocytes and almost all RBCs were in fractions 1 and 2 with lymphocytes. These results are similar to those of other reports which evaluated the same instrument to isolate monocytes from small numbers of PBMC concentrates collected from healthy subjects [7,9,11,20–23], pooled whole blood buffy coat products [8,24] and PBMCs from up to 24 patients with cancer [10,11,16]. …”

Section: Discussionsupporting

confidence: 90%

Counter-flow elutriation of clinical peripheral blood mononuclear cell concentrates for the production of dendritic and T cell therapies

et al. 2014

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IntroductionPeripheral blood mononuclear cells (PBMC) concentrates collected by apheresis are frequently used as starting material for cellular therapies, but the cell of interest must often be isolated prior to initiating manufacturing.Study design and methodsThe results of enriching 59 clinical PBMC concentrates for monocytes or lymphocytes from patients with solid tumors or multiple myeloma using a commercial closed system semi-automated counter-flow elutriation instrument (Elutra, Terumo BCT) were evaluated for quality and consistency. Elutriated monocytes (n = 35) were used to manufacture autologous dendritic cells and elutriated lymphocytes (n = 24) were used manufacture autologous T cell therapies. Elutriated monocytes with >10% neutrophils were subjected to density gradient sedimentation to reduce neutrophil contamination and elutriated lymphocytes to RBC lysis.ResultsElutriation separated the PBMC concentrates into 5 fractions. Almost all of the lymphocytes, platelets and red cells were found in fractions 1 and 2; in contrast, most of the monocytes, 88.6 ± 43.0%, and neutrophils, 74.8 ± 64.3%, were in fraction 5. In addition, elutriation of 6 PBMCs resulted in relatively large quantities of monocytes in fractions 1 or 2. These 6 PBMCs contained greater quantities of monocytes than the other 53 PBMCs. Among fraction 5 isolates 38 of 59 contained >10% neutrophils. High neutrophil content of fraction 5 was associated with greater quantities of neutrophils in the PBMC concentrate. Following density gradient separation the neutrophil counts fell to 3.6 ± 3.4% (all products contained <10% neutrophils). Following red cell lysis of the elutriated lymphocyte fraction the lymphocyte recovery was 86.7 ± 24.0% and 34.3 ± 37.4% of red blood cells remained.ConclusionsElutriation was consistent and effective for isolating monocytes and lymphocytes from PBMC concentrates for manufacturing clinical cell therapies, but further processing is often required.

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Section: Discussionsupporting

confidence: 90%

Counter-flow elutriation of clinical peripheral blood mononuclear cell concentrates for the production of dendritic and T cell therapies

et al. 2014

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“…Since it was first introduced in the 1970s [24–28], counterflow centrifugal elutriation (CCE) has been used extensively in research applications to separate cell products on the basis of size and density. More recently, a clinical elutriation device has been developed (Elutra; GambroBCT, Lakewood, CO, http://www.caridianbct.com) and has been successfully used to isolate monocytes from peripheral blood apheresis products for vaccine applications [29–36] and lymphocytes for adoptive immunotherapy [37]. During elutriation, platelets and red blood cells are efficiently separated from white blood cells with monocytes highly enriched in a single fraction.…”

Section: Introductionmentioning

confidence: 99%

Optimized Processing of Growth Factor Mobilized Peripheral Blood CD34+ Products by Counterflow Centrifugal Elutriation

Tran¹,

Torres-Coronado²,

Gardner³

et al. 2012

Stem Cells Translational Medicine

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Cell separation by counterflow centrifugal elutriation has been described for the preparation of monocytes for vaccine applications, but its use in other current good manufacturing practice (cGMP) operations has been limited. In this study, growth factor-mobilized peripheral blood progenitor cell products were collected from healthy donors and processed by elutriation using a commercial cell washing device. Fractions were collected for each product as per the manufacturer's instructions or using a modified protocol developed in our laboratory. Each fraction was analyzed for cell count, viability, and blood cell differential. Our data demonstrate that, using standard elutriation procedures, >99% of red blood cells and platelets were removed from apheresis products with high recoveries of total white blood cells and enrichment of CD34؉ cells in two of five fractions. With modification of the basic protocol, we were able to collect all of the CD34؉ cells in a single fraction. The CD34-enriched fractions were formulated, labeled with a ferromagnetic antibody to CD34, washed using the Elutra device, and transferred directly to a magnetic bead selection device for further purification. CD34؉ cell purities from the column were extremely high (98.7 ؎ 0.9%), and yields were typical for the device (55.7 ؎ 12.3%). The processes were highly automated and closed from receipt of the apheresis product through formulation of target-enriched cell fractions. Thus, elutriation is a feasible method for the initial manipulations associated with primary blood cell therapy products and supports cGMP and current good tissue practice-compliant cell processing. STEM CELLS TRANSLATIONAL MEDICINE 2012;1:422-429

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Cell Cultures

Bánfalvi

2013

Homeostasis - Tumor - Metastasis

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Clinical-scale elutriation as a means of enriching antigen-presenting cells and manipulating alloreactivity

Cited by 5 publications

References 30 publications

Counter-flow elutriation of clinical peripheral blood mononuclear cell concentrates for the production of dendritic and T cell therapies

Counter-flow elutriation of clinical peripheral blood mononuclear cell concentrates for the production of dendritic and T cell therapies

Optimized Processing of Growth Factor Mobilized Peripheral Blood CD34+ Products by Counterflow Centrifugal Elutriation

Cell Cultures

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