Comparison of platelet aggregability and P-selectin surface expression on platelets isolated by different methods.
Three methods commonly used for isolation of blood platelets from plasma were compared. Platelets were isolated by: 1) a washing method; 2) a method of metrizamide-gradient centrifugation; 3) a modified method of gel-filtration. The last method employed BSA-Sepharose gel instead of routinely used Sepharose gel saturated with BSA. BSA-Sepharose gel was prepared by covalent binding of thermally deactivated BSA to CNBr-activated Sepharose 2B. In contrast to platelets isolated by the other methods, an aggregability of the gel-filtered platelets and control platelets in plasma, both activated with ADP, were comparable. When expression of P-selectin on the surface of freshly isolated platelets was examined, the gel-filtered platelets exhibited the same extent of fluorescence signal as platelets in the citrated blood, whereas platelets isolated by the other methods exhibited twice the extent of the signal. The methods involving the centrifugation process cause a low but a significant platelet activation.
Walkowiak
B
B
Department of Biophysics, Medical University of Lodz, Poland. bogdan01@psk2.am.lodz.pl
Kralisz
U
U
Michalec
L
L
Majewska
E
E
Koziolkiewicz
W
W
Ligocka
A
A
Cierniewski
C S
CS
D000244:Adenosine Diphosphate / Q000494:pharmacology; D001792:Blood Platelets / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000097:blood / Q000502:physiology / Q000648:ultrastructure; D002469:Cell Separation / Q000379:methods; D002498:Centrifugation; D002850:Chromatography, Gel; D006801:Humans; D008722:Methods; D019007:P-Selectin / Q000097:blood* / Q000502:physiology / Q000648:ultrastructure; D010974:Platelet Aggregation / Q000187:drug effects; D015203:Reproducibility of Results; D013917:Thrombin / Q000494:pharmacology
D015815:Cell Adhesion Molecules; D019007:P-Selectin; D000244:Adenosine Diphosphate; D013917:Thrombin; D002118:Calcium