A Simple Method for the Cryopreservation of Human Lymphocytes at −80C

Simon, James D.; Flinton, Linda J.; Albala, Maurice M.

doi:10.1046/j.1537-2995.1977.17177128879.x

Cited by 9 publications

(1 citation statement)

References 27 publications

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“…Cytokine mechanisms in plasma may also be active at higher temperatures and so plasma was also stored at -80°C. Once frozen, samples can be stored in this manner for prolonged periods o f time without damage (Simon 1977).…”

Section: Collection and Storage Of Samplesmentioning

confidence: 99%

The role of the CCR5 Δ32 polymorphism in abdominal aortic aneurysms

Sandford¹,

Bown²,

London³

et al. 2009

Int J Immunogenetics

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All rights reserved INFORMATION TO ALL U SE R S The quality of this reproduction is d ep endent upon the quality of the copy submitted.In the unlikely event that the author did not sen d a com plete manuscript and there are m issing p a g es, th e se will be noted. Also, if material had to be removed, a note will indicate the deletion. Dissertation Publishing UMI U 526187Published by ProQ uest LLC 2013. Copyright in the Dissertation held by the Author. Microform Edition © ProQ uest LLC. All rights reserved. This work is protected against unauthorized copying under Title 17, United States C ode. ProQ uest LLC 789 East Eisenhow er Parkway P.O. Box 1346 Ann Arbor, Ml 48106-1346 ST A TEM EN T O F O R IG IN A LIT YI hereby declare that the work contained in this thesis is entirely my own, except where acknowledged, and has not been submitted for a degree to this or any other University. R.M.Sandford A B ST R A C T The Role o f the CCR5 A 32 Polymorphism in Abdominal Aortic Aneurysms By R.M.SandfordC-C Chemokine receptor 5 (CCR5) is involved in the regulation o f the inflammatory response. Abdominal aortic aneurysms (AAA) may arise as the result o f a chronic inflammatory process which is influenced by a genetic predisposition. The CCR5 gene harbours a 32 base pair deletion (the A32 polymorphism, rs333). The aim o f this study was to investigate the role o f the CCR5 A32 polymorphism in the development of AAA.A case control study was conducted including 285 patients with AAA and 273 control subjects. A blood sample was taken from each individual and DNA extracted. CCR5 genotype was determined using the polymerase chain reaction (PCR). Flow cytometry was used to investigate the biological activity o f the A32 polymorphism.There was no significant difference between the AAA and the control group in relation to the A32 allele frequency (AAA group 10%, control group 12%, P=0.82, chi squared analysis). There was also no significant difference in CCR5 genotype between groups (wild type homozygotes 82% in AAA group vs 77% in control group, heterozygotes 16% in AAA group vs 21% in control group, and A32 homozygotes 2% in both groups, P=0.33, chi squared analysis). The polymorphism was shown to be biologically active with the number o f A32 alleles correlating with cell expression of CCR5 as detected with flow cytometry (P=<0.05).This study demonstrates that the CCR5 A32 is a biologically active genetic polymorphism, however, there is no association between this polymorphism and AAA. Firstly, to Professor Rob Sayers for allowing me the opportunity to undertake this research and also for his supervision and support throughout the project. P U B L IC A T IO N S A R IS IN G FROM T H IS T H E S ISSandford DM Diabetes Mellitus DNAAlso, to Mr M iff Bown for his friendship, patience and encouragement in addition to providing statistical and scientific support. Demographic data 103 Table 7.2: Patient and control subject co-morbidity 105 Table 7.3: Patient and control group medication 106 Table 7.4: Univariate analysis of demog...

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Section: Collection and Storage Of Samplesmentioning

confidence: 99%

The role of the CCR5 Δ32 polymorphism in abdominal aortic aneurysms

Sandford¹,

Bown²,

London³

et al. 2009

Int J Immunogenetics

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Inexpensive Method for Cryopreservation of Human Peripheral Blood Lymphocytes

Assendelft¹,

Jones²,

Stoddard³

et al. 1990

Cryopreservation and Low Temperature Biology in Blood Transfusion

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The maintenance of B-cell and T-cell function in frozen and stored human lymphocytes

et al. 1981

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The objective of this study was to identify and test a convenient means for long-term storage of lymphocytes taken from clinically characterized patients without losing B- or T-cell function. Accordingly, peripheral blood lymphocytes were frozen and stored, and portions of each sample were subsequently assayed for T-cell blastogenic response and B-cell Jerne plaquing at various time intervals after freezing. A comparison of the cell counts of fresh and frozen cultures indicated that cell were recovered after freezing. Furthermore, these cells showed no significant differences in (i) cell viability; (ii) blastogenic response to antigens of Actinomyces maeslandii, Bacteroides melaninogenicus, Fusobacterium nucleatum, and tetanus toxoid; (iii) blastogenic response to phytohemagglutinin and pokeweed mitogen; and (iv) polyclonal B-cell response to pokeweed mitogen as measured by the direct Jerne plaque assay. The retained blastogenic and plaquing responses seen in frozen cultures indicated the maintenance of both T-cell and B-cell function, respectively. This is the first reported demonstration of Jerne plaquing of normal human lymphocytes after freezing. It appears that freezing techniques provide a means for repeating and extending both T- and B-cell assays using frozen stored portions of the same cell samples.

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A Simple Method for the Cryopreservation of Human Lymphocytes at −80C

Cited by 9 publications

References 27 publications

The role of the CCR5 Δ32 polymorphism in abdominal aortic aneurysms

The role of the CCR5 Δ32 polymorphism in abdominal aortic aneurysms

Inexpensive Method for Cryopreservation of Human Peripheral Blood Lymphocytes

The maintenance of B-cell and T-cell function in frozen and stored human lymphocytes

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