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Kordon Kanı Hücrelerinin Saklanmasında Vitrifikasyon Metodu

Vitrification method in banking of cord blood mononuclear cells

Journal Name:

  • Cerrahpaşa Tıp Dergisi

Publication Year:

  • 2009

Key Words:

Keywords (Original Language):

Author NameUniversity of AuthorFaculty of Author
Abstract (2. Language): 
Objectives: The idea of protection of stem cells in a cord blood bank for treatment of various diseases, becomes more widespread. The methods when used during the protection, must be developed for minimizing the risk of cell lost. According to our purpose, we made a comparison by using different protection methods for the cord blood stem cells, during the experiments. Methods: “Density-gradient” method was used for mononuclear cell separation from cord blood, which is taken from voluntary donors. We applied 2 different vitrification method. One of them consist of two steps. Before and after the freezing process, cell viabilities by trypan blue dye method, differences in CD34+, CD45+ and CD34+45+ cells ratio by flow cytometry method and, differences in morphology by light microscope had been determinated. Results: According to our experiment results, CD45+, CD34+ and CD34+/45+ cell ratios significantly decreased after vitrification processes. But there is no significant differences between 2 different vitrification method, we used. In the light microscopy analysis, cell membran structures have damaged, cells have seen like swell and cells come together and have seen attaching. Conclusion: Vitrification methods are usefull for cell banking because of their short, cheap and easy process. We are thinking that our study is helpful for improving new and suitable methods.
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Abstract (Original Language): 
Amaç: Çeşitli hastalıkların tedavisinde uygulayabilmek amacı ile kök hücrelerin saklanarak sürekli bir kök hücre bankasında toplanması fikri giderek yaygın hale gelmektedir. Saklanma sırasında hücre kaybının en aza indirilmesi için kullanılan metotların geliştirilmesi gerekmektedir. Çalışmamızda, kordon kanlarından ayrılan mononükleer hücrelere farklı saklama yöntemleri uygulayarak, yapılan karşılaştırmalar sonucunda hangi yöntemin daha verimli olduğunu belirlemek amaçlanmıştır. Yöntem: Gönüllü vericilerden alınan kordon kanlarından mononükleer hücreler, yoğunluk derecelendirme yöntemi kullanılarak ayrıldı. Biri tek aşamalı diğeri iki aşamalı olarak 2 ayrı vitrifikasyon metodu uygulandı. Vitrifikasyon öncesi ve sonrasında tripan mavisi boyama yöntemi ile hücre canlılığındaki, akım sitometri tekniği kullanılarak CD34+, CD45+ ve CD34+45+ hücre oranlarındaki, ışık mikroskobu kullanarak morfolojide meydana gelen farklılıklar belirlendi. Bulgular: Elde ettiğimiz bulgular sonucunda, dondurma öncesi ve sonrasında CD45+, CD34+, CD34+/45+ hücre oranlarında anlamlı düşüşler gözlendi. İstatistiksel değerlendirmelerde, uygulanan iki farklı vitrifikasyon metodu arasında anlamlı bir fark görülmedi. Yapılan ışık mikroskobi değerlendirmelerinde ise hücrelerin dondurma sonrasında membran yapılarının bozulduğu, hücrelerde şişmeler oluştuğu ve hücre kümeleri meydana geldiği görüldü. Sonuç: Çalışmamızda vitrifikasyon sonrası hücre kayıpları saptansa da kordon kanı hücrelerinin saklanmasında zamandan ve maddi açıdan tasarruf sağlayacak vitrifikasyon tekniklerinin kullanılabilirliğinin sağlanmasının, kordon kanı bankaları ve diğer kök hücre depolama merkezleri açısından da kullanışlı olacağı açıktır.
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  • Turkish

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