Supposedly homogeneous red blood cell (RBC) samples are commonly obtained by “washing” whole blood free of plasma, platelets and white cells with physiologic solutions, a procedure that does not result, however, in sufficient removal of polymorphonuclear neutrophils (PMNs), leading to possible artefactual results. Pure RBC samples can only be obtained by leukodepletion procedures. Proposed here is a version of gelatin zymography adapted to detect matrix-metalloproteinase-9 (MMP-9), selectively expressed by PMNs, in heterogeneous mixtures of RBCs and PMNs, that can reveal contamination at levels as low as 1 PMN/10^6 RBCs.
Application of gelatin zymography for evaluating low-levels of contaminating neutrophils in red blood cell samples
ACHILLI, CESARE;CIANA, ANNARITA;BALDUINI, CESARE;MINETTI, GIAMPAOLO
2011-01-01
Abstract
Supposedly homogeneous red blood cell (RBC) samples are commonly obtained by “washing” whole blood free of plasma, platelets and white cells with physiologic solutions, a procedure that does not result, however, in sufficient removal of polymorphonuclear neutrophils (PMNs), leading to possible artefactual results. Pure RBC samples can only be obtained by leukodepletion procedures. Proposed here is a version of gelatin zymography adapted to detect matrix-metalloproteinase-9 (MMP-9), selectively expressed by PMNs, in heterogeneous mixtures of RBCs and PMNs, that can reveal contamination at levels as low as 1 PMN/10^6 RBCs.File in questo prodotto:
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