Cryogenic preservation of monocytes from human blood and plateletpheresis cellular residues

Hunt, Stephen; Lionetti, Fabian J.; Valeri, C. R.; Callahan, Arthur B.

doi:10.1182/blood.v57.3.592.592

Cited by 20 publications

(2 citation statements)

References 12 publications

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“…Monocytes. Blood monocytes were isohted by density gradient centrifugation followed by counterflow centrifugation ehtriation as described (7). Preparations contained 87 _+ 5% monocytes as determined by light scatter, Wright-Giemsa stain and surface antigen analysis.…”

Section: Methodsmentioning

confidence: 99%

Monocyte attachment to activated human vascular endothelium in vitro is mediated by leukocyte adhesion molecule-1 (L-selectin) under nonstatic conditions.

Spertini

Luscinskas

Gimbrone

et al. 1992

The Journal of Experimental Medicine

140

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SummaryThe receptors that mediate monocyte adhesion to cytokine-stimulated endothelial monolayers were assessed using a nonstatic (rotating) cell-attachment assay. In this system, leukocyte adhesion molecule-1 (LAM-1) (L-sdectin) mediated a major portion (87 _+ 15% at 37~ of monocyte attachment to activated endothelium, mAb blocking of endothelial leukocyte adhesion molecule.1 (41% inhibition), CD18 (36%), and vascular cell adhesion molecule.1 (25%) function had lesser effects on attachment. These results suggest that LAM-1 may serve an important role in monocyte attachment to endothelium at sites of inflammation.

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Section: Methodsmentioning

confidence: 99%

Monocyte attachment to activated human vascular endothelium in vitro is mediated by leukocyte adhesion molecule-1 (L-selectin) under nonstatic conditions.

Spertini

Luscinskas

Gimbrone

et al. 1992

The Journal of Experimental Medicine

140

View full text Add to dashboard Cite

show abstract

“…The main reason is that when a non-permeating cryoprotectant (such as HES) is used in addition to Me 2 SO, HES promotes the water efflux from inside the cell at higher sub-zero temperatures (between −10˚C to −20˚C) when the other solutes are less concentrated compared to when HES is not present; accordingly, HES reduces the amount of intracellular ice formation during plunging into liquid nitrogen [18]. Among different combinations of Me 2 SO and HES, 5% Me 2 SO and 6% HES has been shown to be the most effective combination by previous studies for blood stem cells [34,[40][41][42][43][44][45], human granulocytes [37], bone marrow [46], monocytes [47], canine bone marrow [48], baboon granulocytes [49], and guinea pig granulocytes [50]. Me 2 SO has been reported to be toxic for neurons and astrocytes [51,52] even at very small concentrations.…”

Section: Introductionmentioning

confidence: 99%

Systematic cryopreservation study of cardiac myoblasts in suspension

Ashrafi,

Radisic,

Elliott

2024

PLoS ONE

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H9c2 myoblasts are a cell line derived from embryonic rat heart tissue and demonstrate the ability to differentiate to cardiac myotubes upon reduction of the serum concentration (from 10% to 1%) and addition of all-trans retinoic acid in the growth medium. H9c2 cells are increasingly being used as an easy-to-culture proxy for some functions of cardiomyocytes. The cryobiology of cardiac cells including H9c2 myoblasts has not been studied as extensively as that of some cell types. Consequently, it is important to characterize the cryobiological response and systematically develop well-optimized cryopreservation protocols for H9c2 cells to have optimal and consistent viability and functionality after thaw for high quality studies with this cell type. In this work, an interrupted slow cooling protocol (graded freezing) was applied to characterize H9c2 response throughout the cooling profile. Important factors that affect the cell response were examined, and final protocols that provided the highest post-thaw viability are reported. One protocol uses the common cryoprotectant dimethyl sulfoxide combined with hydroxyethyl starch, which will be suitable for applications in which the presence of dimethyl sulfoxide is not an issue; and the other protocol uses glycerol as a substitute when there is a desire to avoid dimethyl sulfoxide. Both protocols achieved comparable post-thaw viabilities (higher than 80%) based on SYTO 13/GelRed flow cytometry results. H9c2 cells cryopreserved by either protocol showed ability to differentiate to cardiac myotubes comparable to fresh (unfrozen) H9c2 cells, and their differentiation to cardiac myotubes was confirmed with i) change in cell morphology, ii) expression of cardiac marker troponin I, and iii) increase in mitochondrial mass.

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A chemiluminescence assay for erythrophagocytosis

Downing

Templeton

Mitchell

et al. 1990

J. Biolumin. Chemilumin.

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A luminol-dependent chemiluminescence assay for the assessment of the phagocytosis of erythrocytes sensitized with anti-D IgG immunoglobulin by mononuclear leukocytes is described. The mononuclear leukocytes were obtained by apheresis enriched by centrifugation through a density gradient and stored in liquid nitrogen before use. The total reaction mixture, consisting of mononuclear leukocytes-luminol-erythrocytes (either anti-D IgG sensitized or unsensitized controls) was 500 microliters, light detection was by an LKB 1251 luminometer. Peak luminescence was seen between 35-45 minutes, the reaction being exhausted by 120 minutes. Determination of the reproducibility of the assay gave intra- and inter-assay coefficients of variation of 5% and 13% respectively. We found the chemiluminescent response to be affected by the number of erythrocytes used in the assay and by the composition of the medium in which the cells were resuspended, particularly the pH at the initiation of the assay. We also compared the chemiluminescence assay to a microscopic phagocytic assay and found the results virtually identical. However, the former chemiluminescence assay was much easier to perform, marginally more sensitive, less laborious and eliminated any possibility of subjective error.

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Cryogenic preservation of monocytes from human blood and plateletpheresis cellular residues

Cited by 20 publications

References 12 publications

Monocyte attachment to activated human vascular endothelium in vitro is mediated by leukocyte adhesion molecule-1 (L-selectin) under nonstatic conditions.

Monocyte attachment to activated human vascular endothelium in vitro is mediated by leukocyte adhesion molecule-1 (L-selectin) under nonstatic conditions.

Systematic cryopreservation study of cardiac myoblasts in suspension

A chemiluminescence assay for erythrophagocytosis

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