Effect of ex vivo storage and Cyto-Chex on the expression of P-selectin glycoprotein ligand-1 (PSGL-1) on human peripheral leukocytes.
P-selectin glycoprotein ligand-1 (PSGL-1) is expressed on leukocytes and its expression is elevated in asthma and chronic obstructive pulmonary disease. This study assessed whether the cell stabilisation solution Cyto-Chex is able to preserve such quantitative differences for delayed testing. Peripheral blood was mixed with Cyto-Chex and stored refrigerated or at ambient temperature. Aliquots were tested by flow cytometry 2 to 168 h after collection for PSGL-1, CD3, CD4, CD8, and CD16 expression and marker-positive cell counts. Compared with controls a marked reduction of staining intensities is seen at all time points for all markers, irrespective of the storage temperature. This loss of bright staining did not or only moderately decrease marker-positive cell counts except for PSGL-1+ lymphocytes which declined in parallel with staining intensity. These findings indicate that Cyto-Chex is not able to preserve the expression or affinity to antibodies of surface markers to allow delayed determination of quantitative differences or detection of weakly staining cells. Immunophenotyping is mostly possible for 7 days after collection, however, this has to be tested for each marker and cell type of interest.
Schumacher
Axel
A
Department of Internal Medicine/Infectiology and Pneumology, Division of Pneumology, Charité-Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany. axel.schumacher@charite.de
D001793:Blood Preservation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007962:Leukocytes / Q000378:metabolism*; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D019188:Organ Preservation Solutions; D013696:Temperature
D008562:Membrane Glycoproteins; D019188:Organ Preservation Solutions; C084882:P-selectin ligand protein