Lymphocyte subpopulations do not alter during blood storage at 4°C

Lambalgen, R. Van; Meurs, G.J.E. Van

doi:10.1016/0022-1759(85)90162-0

Cited by 15 publications

(9 citation statements)

References 2 publications

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“…It is not known whether there were negative effects from storing or shipping at 4°C. Our data show that there is better viability, cell yield, and function when cells are shipped at room temperature (22°C) or 30°C than at 15°C, and it is generally accepted that storage of whole blood at 4°C negatively impacts cell viability [5], function [29], and population recovery [6,7,43,44]. Acknowledging the range of data in the literature, in a separate study, we are also evaluating the function of PBMC processed the same day (< 8 h) or after overnight shipping or storage (manuscript in preparation).…”

Section: Discussionmentioning

confidence: 93%

Shipping blood to a central laboratory in multicenter clinical trials: effect of ambient temperature on specimen temperature, and effects of temperature on mononuclear cell yield, viability and immunologic function

Olson

Smolkin

Farris³

et al. 2011

J Transl Med

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BackgroundClinical trials of immunologic therapies provide opportunities to study the cellular and molecular effects of those therapies and may permit identification of biomarkers of response. When the trials are performed at multiple centers, transport and storage of clinical specimens become important variables that may affect lymphocyte viability and function in blood and tissue specimens. The effect of temperature during storage and shipment of peripheral blood on subsequent processing, recovery, and function of lymphocytes is understudied and represents the focus of this study.MethodsPeripheral blood samples (n = 285) from patients enrolled in 2 clinical trials of a melanoma vaccine were shipped from clinical centers 250 or 1100 miles to a central laboratory at the sponsoring institution. The yield of peripheral blood mononuclear cells (PBMC) collected before and after cryostorage was correlated with temperatures encountered during shipment. Also, to simulate shipping of whole blood, heparinized blood from healthy donors was collected and stored at 15°C, 22°C, 30°C, or 40°C, for varied intervals before isolation of PBMC. Specimen integrity was assessed by measures of yield, recovery, viability, and function of isolated lymphocytes. Several packaging systems were also evaluated during simulated shipping for the ability to maintain the internal temperature in adverse temperatures over time.ResultsBlood specimen containers experienced temperatures during shipment ranging from -1 to 35°C. Exposure to temperatures above room temperature (22°C) resulted in greater yields of PBMC. Reduced cell recovery following cryo-preservation as well as decreased viability and immune function were observed in specimens exposed to 15°C or 40°C for greater than 8 hours when compared to storage at 22°C. There was a trend toward improved preservation of blood specimen integrity stored at 30°C prior to processing for all time points tested. Internal temperatures of blood shipping containers were maintained longer in an acceptable range when warm packs were included.ConclusionsBlood packages shipped overnight by commercial carrier may encounter extreme seasonal temperatures. Therefore, considerations in the design of shipping containers should include protecting against extreme ambient temperature deviations and maintaining specimen temperature above 22°C or preferably near 30°C.

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Section: Discussionmentioning

confidence: 93%

Shipping blood to a central laboratory in multicenter clinical trials: effect of ambient temperature on specimen temperature, and effects of temperature on mononuclear cell yield, viability and immunologic function

Olson

Smolkin

Farris³

et al. 2011

J Transl Med

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“…Specimen storage has also been evaluated as a technical variable in the immunophenotyping of normal peripheral blood and appears to be closely linked with the consideration of which anticoagulant is most appropriate for routine use (33)(34)(35)(36)(37)(38)(39)(40). Several studies have indicated that poor immunophenotyping results are obtained by prolonged storage of peripheral blood in EDTA (35,38).…”

Section: Peripheral Bloodmentioning

confidence: 99%

Quality control in flow cytometry for diagnostic pathology: II. A conspectus of reference ranges for lymphocyte immunophenotyping

McCoy

Overton

1994

Cytometry

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Immunophenotyping, as many other clinical assays, is interpreted only in the context of reference values obtained from healthy control individuals. While the use of these reference values, or ranges, has been commonplace in the clinical flow cytometry laboratory for well over a decade, there has been little consensus in standardizing how these values should be obtained, analyzed, or expressed. This report reviews the variables to be considered in establishing reference ranges and statistical methods which can be used. Additionally, examples are given of previously published reference ranges for a variety of specimens often submitted for immunophenotyping. D

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“…Por outro lado, muitos autores relataram mudanças resultantes do armazenamento das amostras de sangue (Weiblen et al 1984, Ekong et al 1992. Muitas dessas mudanças podem ser explicadas, pelo menos em parte, pela contaminação do "gate" de linfócitos por neutrófilos (Van Lambalgen & Van Meurs 1985). Lloyd et al (1995) não detectaram mudanças em amostras de sangue ovino armazenado em temperatura ambiente por 48 horas.…”

Section: Discussionunclassified

Quantificação de subpopulações linfocitárias no sangue periférico de cães sadios adultos

et al. 2017

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RESUMO: A citometria de fluxo vem se firmando como uma ferramenta útil na prática médico-veterinária, particularmente na clínica de pequenos animais. Tal conhecimento tem ensejado o estabelecimento de valores fisiológicos para diferentes subpopulações linfocitárias, indispensáveis à compreensão da dinâmica da celularidade linfóide, em diversas situações patológicas. Assim sendo, o presente ensaio teve como objetivo imunomarcar, por intermédio da técnica citométrica, as subpopulações linfocitárias CD5+CD4+, CD5+CD8+ e CD21+, em quatro diferentes raças de cães domésticos e sadios, de tal forma a agregar informações sobre o perfil imunológico das diferentes raças. Foram utilizados 40 cães adultos (2-7 anos), machos e fêmeas, das raças Beagle (G1, n=10), Golden Retriever (G2, n=10), Bulldog Inglês (G3, n=10) e sem raça definida (SRD) (G4, n=10). As colheitas de sangue foram realizadas por venipunção jugular, utilizando-se sistema de frascos a vácuo (K2-EDTA). O hemograma e o processamento das amostras para citometria de fluxo foram realizados num prazo máximo de 24 horas após a colheita do sangue. As amostras foram analisadas no citofluorômetro FACSCANTO (Becton Dickinson, San Jose, CA, USA). Utilizou-se o programa FACSDiva (Becton Dickinson, San Jose, CA, USA), para identificar e quantificar as células CD5+CD4+, CD5+CD8+ e CD21+, que forneceu o histograma e respectiva tabela com a quantidade de células detectadas pela imunofenotipagem. Os dados obtidos para contagem de linfócitos T auxiliares, T citotóxicos/supressores e linfócitos B foram tabulados e submetidos a Análise de Variância pelo teste F. O teste de Tukey a 5% de probabilidade foi utilizado para comparação das médias entre as diferentes raças de cães. Os valores médios de contagens de células CD5+CD8+ no sangue periférico do G1, G2, G3 e G4 foram de 155, 206, 544 e 503 células/μL, respectivamente. Os valores médios de contagens de células CD5+CD4+ no sangue periférico do G1, G2, G3 e G4 foram de 746, 642, 1101 e 855 células/μL, respectivamente. Os valores médios de contagens de células CD21+ no sangue periférico do G1, G2, G3 e G4 foram de 171, 299, 494 e 403 células/μL, respectivamente. Assim sendo, o número médio de células obtido para a subpopulação de linfócitos T citotóxicos foi significativamente maior (p<0,05) nos Bulldogs Ingleses e cães SRD, comparativamente aqueles encontrados nos grupos de Beagles e Golden Retrievers. Ademais, observou-se que o número médio de células obtido para a subpopulação de linfócitos B foi significavamente maior (p<0,05) nos Bulldogs Ingleses, quando comparado àquele dos Beagles.

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Lymphocyte subpopulations do not alter during blood storage at 4°C

Cited by 15 publications

References 2 publications

Shipping blood to a central laboratory in multicenter clinical trials: effect of ambient temperature on specimen temperature, and effects of temperature on mononuclear cell yield, viability and immunologic function

Shipping blood to a central laboratory in multicenter clinical trials: effect of ambient temperature on specimen temperature, and effects of temperature on mononuclear cell yield, viability and immunologic function

Quality control in flow cytometry for diagnostic pathology: II. A conspectus of reference ranges for lymphocyte immunophenotyping

Quantificação de subpopulações linfocitárias no sangue periférico de cães sadios adultos

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