Red cell investigations: Art and artefacts

Minetti, Giampaolo; Egée, Stéphane; Mörsdorf, Daniel; Steffen, Patrick R.; Makhro, Asya; Achilli, Cesare; Ciana, Annarita; Wang, Jue; Bouyer, Guillaume; Bernhardt, Ingolf; Wagner, Christian; Thomas, Serge; Bogdanova, Anna; Kaestner, Lars

doi:10.1016/j.blre.2013.02.002

Cited by 75 publications

(68 citation statements)

References 82 publications

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Section: Experiments On Erythrocyte Adhesionmentioning

confidence: 99%

“…Experiments on aggregation of red cells have been fundamental to the understanding of blood types and many complex diseases including infections due to malaria and virus parasites [23][24][25]. But the problems of purifying and standardizing red cells remain significant [24]. In this study, cells were prepared from three species, human blood from North Staffordshire Hospital, fresh horse blood in EDTA and fresh rat blood from Central Animal Pathology Ltd. Each blood sample was washed six to seven times in phosphate buffered saline to remove the non-red cell components, before suspending in physiological saline solution, then examined by both optical and Coulter tests.…”

Section: Experiments On Erythrocyte Adhesionmentioning

confidence: 99%

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van der Waals forces influencing adhesion of cells

Kendall

Roberts

2015

Phil. Trans. R. Soc. B

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Adhesion molecules, often thought to be acting by a 'lock and key' mechanism, have been thought to control the adhesion of cells. While there is no doubt that a coating of adhesion molecules such as fibronectin on a surface affects cell adhesion, this paper aims to show that such surface contamination is only one factor in the equation. Starting from the baseline idea that van der Waals force is a ubiquitous attraction between all molecules, and thereby must contribute to cell adhesion, it is clear that effects from geometry, elasticity and surface molecules must all add on to the basic cell attractive force. These effects of geometry, elasticity and surface molecules are analysed. The adhesion force measured between macroscopic polymer spheres was found to be strongest when the surfaces were absolutely smooth and clean, with no projecting protruberances. Values of the measured surface energy were then about 35 mJ m 22 , as expected for van der Waals attractions between the non-polar molecules. Surface projections such as abrasion roughness or dust reduced the molecular adhesion substantially. Water cut the measured surface energy to 3.4 mJ m 22 . Surface active molecules lowered the adhesion still further to less than 0.3 mJ m 22 . These observations do not support the lock and key concept.

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Section: Experiments On Erythrocyte Adhesionmentioning

confidence: 99%

Section: Experiments On Erythrocyte Adhesionmentioning

confidence: 99%

van der Waals forces influencing adhesion of cells

Kendall

Roberts

2015

Phil. Trans. R. Soc. B

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“…This process was suggested to produce pure erythrocyte preparations without the remaining leucocytes and platelets. While this protocol is well established and widely used in blood cell investigations (see, for instance 74 , for a recent review), the ghost solution produced by this protocol did not result in well developed multi-lamellar membrane stacks when applied on silicon wafers. Cellulose particles were observed under the microscope in the solution after passing through the filter (shown in the Supplementary Material in Fig.…”

Section: Preparation Of Ghosts the Preparation Of Rbc Ghosts Was Firmentioning

confidence: 99%

The Molecular Structure of Human Red Blood Cell Membranes From Highly Oriented, Solid Supported Multi-Lamellar Membranes

Himbert

Alsop

Rheinstädter

2018

Biophysical Journal

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We prepared highly oriented, multi-lamellar stacks of human red blood cell (RBC) membranes applied on silicon wafers. RBC ghosts were prepared by hemolysis and applied onto functionalized silicon chips and annealed into multi-lamellar RBC membranes. High resolution X-ray diffraction was used to determine the molecular structure of the stacked membranes. We present direct experimental evidence that these RBC membranes consist of nanometer sized domains of integral coiled-coil peptides, as well as liquid ordered (l o ) and liquid disordered (l d ) lipids. Lamellar spacings, membrane and hydration water layer thicknesses, areas per lipid tail and domain sizes were determined. The common drug aspirin was added to the RBC membranes and found to interact with RBC membranes and preferably partition in the head group region of the l o domain leading to a fluidification of the membranes, i.e., a thinning of the bilayers and an increase in lipid tail spacing. Our results further support current models of RBC membranes as patchy structures and provide unprecedented structural details of the molecular organization in the different domains.The presence of pale cells with no internal content in a blood smear is typically indicative of a disease. These cells are produced by hemolysis and have been named red blood cell (RBC) ghosts based on their appearance under the microscope. RBC ghosts can be prepared artificially [1][2][3] . The first published protocol in 1963 by Dodge, Mitchell and Hanahan describes the extraction of the cell membrane from RBCs through hemolysis and was an essential step in the development of membrane proteomics and lipidomics 4,5 . While there is detailed knowledge of the composition of RBC membranes, information about the molecular organization of these components in the actual membranes is scarce 6 . This is, in particular, a consequence of the lack of suitable experimental techniques. The disordered, patchy and highly dynamic state of biological materials impedes, for instance, the use of high-resolution diffraction techniques as in protein crystallography. To overcome this constraint, we prepared highly oriented stacks of RBC membranes on silicon wafers and studied their molecular properties in-vitro using high resolution X-ray diffraction. The results present evidence for nanometer-sized domains of coiled-coils peptides, as well as liquid ordered (l o ) and liquid disordered (l d ) lipid patches, and give a detailed picture of the molecular organization in these domains.When present in the body, aspirin (acetylsalicylic acid, ASA) and its metabolites interact with the cyclo-oxygenase (COX) pathway. The inhibition of both COX isoforms, COX-1 and COX-2, by higher dose aspirin is believed to lead to analgesic and anti-inflammatory effects, while lower doses, sufficient to inhibit COX-1 activity, lead to anti-platelet activity 7,8 . There is recent evidence that membrane composition and fluidity play an important role in platelet cell function [9][10][11] , possibly related to the formation of rafts 12 .

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“…A number of cell lines cells including HEK293, CHO, LTK, RBL, SHSY5Y, GH4C1, ND7-23 and jurkat are routinely used on the Patchliner with equally high success rates. In addition, currents from primary cells such as rat cortical astrocytes, 20 human fibroblast-like synioviocytes, 20 red blood cells 21 and vascular smooth muscle cells 22 have been successfully recorded. Cardiomyocytes derived from pluripotent stem cells (iPS) are receiving increasing attention as a potential model for safety screening and toxicology for early screening of lead compounds.…”

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confidence: 99%