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Red blood cell phenotype fidelity following glycerol cryopreservation optimized for research purposes.

dc.contributor.author McMahon, Timothy
dc.contributor.author Palmer, Gregory
dc.contributor.author Dosier, Laura
dc.contributor.author Rogers, Stephen C
dc.contributor.author Zhu, Hongmei
dc.contributor.author Timm, David
dc.contributor.author Zhang, Hengtao
dc.contributor.author Herbert, Joseph
dc.contributor.author Atallah, Jacqueline
dc.contributor.author Cook, Asa
dc.contributor.author Ernst, Melanie
dc.contributor.author Prakash, Jaya
dc.contributor.author Terng, Mark
dc.contributor.author Towfighi, Parhom
dc.contributor.author Doctor, Reid
dc.contributor.author Said, Ahmed
dc.contributor.author Joens, Matthew S
dc.contributor.author Fitzpatrick, James AJ
dc.contributor.author Hanna, Gabi
dc.contributor.author Lin, Xue
dc.contributor.author Reisz, Julie A
dc.contributor.author Nemkov, Travis
dc.contributor.author D'Alessandro, Angelo
dc.contributor.author Doctor, Allan
dc.date.accessioned 2019-07-12T20:17:32Z
dc.date.available 2019-07-12T20:17:32Z
dc.date.issued 2018-01
dc.identifier PONE-D-18-25310
dc.identifier.issn 1932-6203
dc.identifier.issn 1932-6203
dc.identifier.uri https://hdl.handle.net/10161/19098
dc.description.abstract Intact red blood cells (RBCs) are required for phenotypic analyses. In order to allow separation (time and location) between subject encounter and sample analysis, we developed a research-specific RBC cryopreservation protocol and assessed its impact on data fidelity for key biochemical and physiological assays. RBCs drawn from healthy volunteers were aliquotted for immediate analysis or following glycerol-based cryopreservation, thawing, and deglycerolization. RBC phenotype was assessed by (1) scanning electron microscopy (SEM) imaging and standard morphometric RBC indices, (2) osmotic fragility, (3) deformability, (4) endothelial adhesion, (5) oxygen (O2) affinity, (6) ability to regulate hypoxic vasodilation, (7) nitric oxide (NO) content, (8) metabolomic phenotyping (at steady state, tracing with [1,2,3-13C3]glucose ± oxidative challenge with superoxide thermal source; SOTS-1), as well as in vivo quantification (following human to mouse RBC xenotransfusion) of (9) blood oxygenation content mapping and flow dynamics (velocity and adhesion). Our revised glycerolization protocol (40% v/v final) resulted in >98.5% RBC recovery following freezing (-80°C) and thawing (37°C), with no difference compared to the standard reported method (40% w/v final). Full deglycerolization (>99.9% glycerol removal) of 40% v/v final samples resulted in total cumulative lysis of ~8%, compared to ~12-15% with the standard method. The post cryopreservation/deglycerolization RBC phenotype was indistinguishable from that for fresh RBCs with regard to physical RBC parameters (morphology, volume, and density), osmotic fragility, deformability, endothelial adhesivity, O2 affinity, vasoregulation, metabolomics, and flow dynamics. These results indicate that RBC cryopreservation/deglycerolization in 40% v/v glycerol final does not significantly impact RBC phenotype (compared to fresh cells).
dc.language eng
dc.publisher Public Library of Science (PLoS)
dc.relation.ispartof PloS one
dc.relation.isversionof 10.1371/journal.pone.0209201
dc.subject Erythrocytes
dc.subject Animals
dc.subject Humans
dc.subject Mice
dc.subject Mice, Nude
dc.subject Glycerol
dc.subject Hemoglobins
dc.subject Cryoprotective Agents
dc.subject Microscopy, Electron, Scanning
dc.subject Erythrocyte Indices
dc.subject Osmotic Fragility
dc.subject Cryopreservation
dc.subject Blood Preservation
dc.subject Erythrocyte Transfusion
dc.subject Transplantation, Heterologous
dc.subject Cell Adhesion
dc.subject Erythrocyte Deformability
dc.subject Phenotype
dc.subject Metabolome
dc.subject Healthy Volunteers
dc.title Red blood cell phenotype fidelity following glycerol cryopreservation optimized for research purposes.
dc.type Journal article
dc.date.updated 2019-07-12T20:17:31Z
pubs.begin-page e0209201
pubs.issue 12
pubs.organisational-group School of Medicine
pubs.organisational-group Duke
pubs.organisational-group Medicine, Pulmonary, Allergy, and Critical Care Medicine
pubs.organisational-group Medicine
pubs.organisational-group Clinical Science Departments
pubs.organisational-group Pediatrics, Pulmonary and Sleep Medicine
pubs.organisational-group Pediatrics
pubs.organisational-group Duke Cancer Institute
pubs.organisational-group Institutes and Centers
pubs.organisational-group Radiation Oncology
pubs.publication-status Published
pubs.volume 13
duke.contributor.orcid Palmer, Gregory|0000-0003-2955-8297


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