Publication: İn vitro şartlarda bekletilen trombositlerde zamana bağlı trombosit protein değişimlerinin elektroforez yöntemiyle incelenmesi
Abstract
Hemostazın önemli hücrelerinden olan trombositlerin kalitatif ve kantitatif bozukluklarında gelişen hemorajik diatezin klinik kontrolünde, trombosit transfüzyonundan yararlanılmaktadır. Bu trombositler trombosit sayısı normal sağlıklı donörlerden elde edilirler, uygulanılan işleme bağlı olarak da farklı sayılarda olurlar. Trombosit elde etmek için en etkili işlem aferez yöntemidir. Kullanılan torbanın özelliğine bağlı olarak da alınmasından sonra bir gün ya da 5 gün kadar sonra 20-24°C’ de özel bir çalkalayıcıda kullanıma hazır olarak bekletilebilirler. Bu çalışmada sağlıklı bireylerden aferez yöntemiyle elde edilen trombositler ajitatörde in vitro şartlarda 9 gün 20–24°C’ de bekletildi. 1., 2., 3., 4., 5. ve 9. günlerde trombosit numuneleri alınarak çöktürüldü ve tamponla yıkandı. Dört kez dondurulup eritilerek patlatıldı. Santrifigasyon işleminden sonra üst faz elde edildi. Lowry metodu ile protein tayini yapıldı ve protein değişimini izlemek için SDS-PAGE (Sodyum dodesil sülfat poliakrilamid jel elektroforezi) uygulandı. Beş gün boyunca in vitro şartlarda bekletilen trombositlerde SDS-PAGE sonucunda protein bantlarında herhangi bir değişiklik görülmedi. INVESTIGATING THE TIME DEPENDENT PLATELET PROTEIN ALTERATIONS IN THE PLATELETS WHICH HAVE BEEN KEPT UNDER IN VITRO CONDITIONS BY ELECTROPHORESIS.
Platelets are one of the most important cells of homeostasis. Platelet transfusion has been utilized in clinical control of the hemorrhagic diathesis caused by qualitative and quantitative disorders of platelets. These platelets are obtained from healthy donors with normal platelet count differed depending on the method used. The most effective method used to obtain platelets is the aphaeresis method. After obtaining, the cells can be kept in a special shaker under 20-24°C for 1 to 5 days. In this study, the platelets obtained from healthy donors by aphaeresis method were kept in an agitator under in-vitro conditions for nine days at 20-24°C. The samples were taken on 1st, 2nd, 3rd, 4th, 5th and 9th days. Platelets were isolated and washed with Tris buffer. Platelets were frozen and thawed 4 times. After centrifugation, the supernatant was obtained. Then protein concentration was determined with the Lowry method and SDS-PAGE (Sodium dodecyl sulfate polyacrylamide gel electrophoresis) was applied to examine the protein alterations. No alternation in the number of protein bands has been observed in SDS-PAGE in 5 days incubation.
Platelets are one of the most important cells of homeostasis. Platelet transfusion has been utilized in clinical control of the hemorrhagic diathesis caused by qualitative and quantitative disorders of platelets. These platelets are obtained from healthy donors with normal platelet count differed depending on the method used. The most effective method used to obtain platelets is the aphaeresis method. After obtaining, the cells can be kept in a special shaker under 20-24°C for 1 to 5 days. In this study, the platelets obtained from healthy donors by aphaeresis method were kept in an agitator under in-vitro conditions for nine days at 20-24°C. The samples were taken on 1st, 2nd, 3rd, 4th, 5th and 9th days. Platelets were isolated and washed with Tris buffer. Platelets were frozen and thawed 4 times. After centrifugation, the supernatant was obtained. Then protein concentration was determined with the Lowry method and SDS-PAGE (Sodium dodecyl sulfate polyacrylamide gel electrophoresis) was applied to examine the protein alterations. No alternation in the number of protein bands has been observed in SDS-PAGE in 5 days incubation.