Are platelets activated after a rapid, one-step density gradient centrifugation? Evidence from flow cytometric...

Area of Science:

• Hematology
• Immunology
• Biochemistry

Background:

• Platelets are crucial for hemostasis and immune response.
• Platelet activation leads to surface marker changes, such as CD63 expression.
• Accurate platelet analysis requires isolation methods that preserve functional integrity.

Purpose of the Study:

• To describe a rapid, one-step density gradient centrifugation method for platelet preparation.
• To evaluate the method's ability to prevent artificial platelet activation.
• To enable reliable immunofluorescence staining of platelet surface markers like CD63.

Main Methods:

• Whole blood from healthy donors and patients with pregnancy-induced hypertension (PIH) was used.
• A one-step density gradient centrifugation technique utilizing OptiPrep was employed for platelet isolation.
• Direct immunofluorescence staining was performed on isolated platelets to detect CD63 expression.

Main Results:

• The described method efficiently prepares platelets from whole blood.
• This technique prevents artificial platelet activation caused by sample manipulation.
• The isolated platelets are suitable for direct immunofluorescence staining and flow cytometry analysis.

Conclusions:

• The one-step density gradient centrifugation method is a reliable technique for platelet isolation.
• This method preserves platelet functional parameters, avoiding artifactual activation.
• It is advantageous for flow cytometry analysis of platelet surface markers, including CD63.