Red blood cell metabolism, preservation, and oxygen delivery

Hess, John R.; Solheim, Bjarte G.

doi:10.1002/9781119013020.ch09

Cited by 6 publications

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“…The RBC numbers collected for the six morphology subclasses were multiplied by fractional weights, which were slightly modified from the conventional MI calculation (4,24) to align with the RBC-MIST's segregation limitation (Section RBC-MIST): The smears were stained with a Hema 3 stain set (Fisher Scientific, Waltham, MA) which is comparable to a Wright-Giemsa staining technique.…”

Section: Lm Rbc Morphology Assaymentioning

confidence: 99%

“…Vesicles budding from the membrane of a biconcave disc (discocyte) produce a crenated appearance around the cell (echinocyte). Eventually, the loss of membrane integrity causes an RBC to morph into a spherical body and is termed a spheroechinocyte (SEC) (4). Eventually, the loss of membrane integrity causes an RBC to morph into a spherical body and is termed a spheroechinocyte (SEC) (4).…”

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confidence: 99%

“…This updated version segregates images into additional RBC morphology subclasses, further aligning these classes with conventional light microscopy-based RBC morphology characterization. RBC morphology nomenclature with associated descriptions and acronyms (4,(19)(20)(21)(22) During unit sampling, RCC units were removed from refrigeration for durations that were within the recommended exposure limit (30). Additionally, the relative percentage of SECs was monitored over the course of RCC unit storage.…”

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confidence: 99%

“…Continual membrane degradation results in an increase in membrane crenations, causing an RBC to lose a greater portion of its membrane support structures (7). Eventually, the loss of membrane integrity causes an RBC to morph into a spherical body and is termed a spheroechinocyte (SEC) (4). The latter morphology transformation is irreversible and results in a distinct lack of deformability when compared to discocytes and echinocytes (8)(9)(10).…”

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“…While recent publications have found no causal link between the storage age of transfused RBCs and mortality rates (14,15), debate still exists on the relationship between storage duration and post-transfusion morbidities (16,17). Regardless of the potential clinical implications, there is widespread agreement that: (1) RBC storage lesions result in an increase of SECs during in vitro storage and (2) this morphology class is less effective than discocytes and echinocytes in delivering oxygen (4). Hence, in the context of augmented oxygen perfusion, RCC possessing a relatively larger percentage of SECs can be considered as lower in quality (18).…”

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Label‐Free Analysis of Red Blood Cell Storage Lesions Using Imaging Flow Cytometry

et al. 2019

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Deleterious changes, collectively termed as storage lesions, alter the characteristics of red blood cell (RBC) morphology during in vitro storage. Due to gradual loss of cellular membrane, RBCs lose their original biconcave disk shape and begin a process of spherical deformation that is characterized by well-defined morphological criteria. At the spheroechinocyte stage, the structure of RBC is irreversibly damaged and lacks the elasticity necessary to efficiently deliver oxygen. Quantifying the prevalence of spheroechinocytes could provide an important morphological measure of the quality of stored blood products. Unlike the conventional RBC morphology characterization assay involving light microscopy, we introduce a label-free assay using imaging flow cytometry (IFC). The technique captures 100,000 images of a sample and calculates a relative measure of spheroechinocyte population in a fraction of the time required by the conventional method. A comparative method study, measuring a morphological index for 11 RCC units through storage, found that the two techniques measured similar trends with IFC reporting the metric at an average of 3.9% higher. We monitored 18 RCC units between Weeks 1 and 6 of storage and found that the spheroechinocyte population increased by an average of 26.2%. The large (3.5-64.1%) variation between the units' spheroechinocyte population percentage at Week 1 suggests a possible dependence of blood product quality on donor characteristics. Given our method's ability to rapidly monitor large samples and refine morphological characterization beyond conventional methods, we believe our technique offers good potential for studying the underlying relationships between RBC morphology and blood storage lesions.Key terms deformability; erythrocyte; imaging flow cytometry; red blood cell morphology; storage lesion CURRENTLY, blood banks and hospital laboratories are devoid of convenient, label-free techniques to measure the morphological distribution of red blood cells (RBCs) stored in vitro. Parameters such as membrane shape and deformability are critical for understanding an RBC's functional efficacy as an oxygen transporter (1). Microscopy-related assays, considered the gold standard for RBC morphology analysis, stain and smear samples prior to manual characterization. High-throughput assays that can measure RBCs within environments closely resembling in vivo conditions could obtain a more insightful assessment of the morphology distribution.RBCs are stored in vitro primarily to enhance the oxygen carrying capacity of vulnerable patients via transfusions (2). Blood banks extract RBCs from donated blood using customized separation techniques and suspend the cells in a preservative fluid to increase its shelf-life (3). During storage of this red cell concentrate (RCC), typically set at a maximum of 6 weeks, RBCs undergo a sequence of significant biochemical and biomechanical changes, which have been collectively termed hypothermic storage lesions (4,5). RBC storage lesions cause the formation of m...

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Section: Lm Rbc Morphology Assaymentioning

confidence: 99%

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Label‐Free Analysis of Red Blood Cell Storage Lesions Using Imaging Flow Cytometry

et al. 2019

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Objective assessment of stored blood quality by deep learning

Doan

Sebastian

Caicedo

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Stored red blood cells (RBCs) are needed for life-saving blood transfusions, but they undergo continuous degradation. RBC storage lesions are often assessed by microscopic examination or biochemical and biophysical assays, which are complex, time-consuming, and destructive to fragile cells. Here we demonstrate the use of label-free imaging flow cytometry and deep learning to characterize RBC lesions. Using brightfield images, a trained neural network achieved 76.7% agreement with experts in classifying seven clinically relevant RBC morphologies associated with storage lesions, comparable to 82.5% agreement between different experts. Given that human observation and classification may not optimally discern RBC quality, we went further and eliminated subjective human annotation in the training step by training a weakly supervised neural network using only storage duration times. The feature space extracted by this network revealed a chronological progression of morphological changes that better predicted blood quality, as measured by physiological hemolytic assay readouts, than the conventional expert-assessed morphology classification system. With further training and clinical testing across multiple sites, protocols, and instruments, deep learning and label-free imaging flow cytometry might be used to routinely and objectively assess RBC storage lesions. This would automate a complex protocol, minimize laboratory sample handling and preparation, and reduce the impact of procedural errors and discrepancies between facilities and blood donors. The chronology-based machine-learning approach may also improve upon humans’ assessment of morphological changes in other biomedically important progressions, such as differentiation and metastasis.

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Scaling of statically derived osteocyte lacunae in extant birds: implications for palaeophysiological reconstruction

Grunmeier

D’Emic

2019

Biol. Lett.

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Osteocytes are mature versions of osteoblasts, bone-forming cells that develop in two ways: via 'static' osteogenesis, differentiating and ossifying tissue in situ to form a scaffold upon which other bone can form, or 'dynamic' osteogenesis, migrating to infill or lay down bone around neurovasculature. A previous study regressed the volume of osteocyte lacunae derived from dynamic osteogenesis (DO) of a broad sample of extant bird species against body mass, the growth rate constant (k), mass-specific metabolic rate, genome size, and erythrocyte size. There were significant relationships with body mass, growth rate, metabolic rate, and genome size, with the latter being the strongest. Using the same avian histological dataset, we measured over 3800 osteocyte lacunar axes derived from static osteogenesis (SO) in order to look for differences in the strength of formfunction relationships inferred for DO-derived lacunae at the cellular and tissue levels. The relationship between osteocyte lacunar volume and body mass was stronger when measuring SO lacunae, whereas relationships between osteocyte lacunar volume versus growth rate and basal metabolic rate disappeared. The relationship between osteocyte lacuna volume and genome size remained significant and moderately strong when measuring SO lacunae, whereas osteocyte lacuna volume was still unrelated to erythrocyte size. Our results indicate that growth and metabolic rate signals are contained in avian DO but not SO osteocyte lacunae, suggesting that the former should be used in estimating these parameters in extinct animals.

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Red blood cell metabolism, preservation, and oxygen delivery

Cited by 6 publications

References 0 publications

Label‐Free Analysis of Red Blood Cell Storage Lesions Using Imaging Flow Cytometry

Label‐Free Analysis of Red Blood Cell Storage Lesions Using Imaging Flow Cytometry

Objective assessment of stored blood quality by deep learning

Scaling of statically derived osteocyte lacunae in extant birds: implications for palaeophysiological reconstruction

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