Kitosan Kaplı Yüzeylerde Kollajen, Vitronektin ve Jelatinin Üç Boyutlu HEK293 Hücre Kültürüne ve Hücresel Kollajen Salınımına Olan Etkileri

Öz

Amaç: Son zamanlarda in vivo ve klinik çalışmalara uyarlanmasının daha kolay olması nedeniyle üç boyutlu kültür oluşturulması oldukça önem kazanmıştır. Üç boyutlu kültür çalışmaları açısından hem maliyeti hem de farklı hücrelerde uygulanabilmesi nedeniyle kitosan materyali ön plana çıkmaktadır. Projemizde hücreler arası matriks elemanlarının kitosan yüzeylere kaplanmasının, hücresel sferoid oluşumuna ve Hücrelerarası Matriks Elemenları HME salınımına olan etkileri araştırılmıştır.Gereç ve Yöntemler: Bu çalışmada, HEK293 insan embriyonik böbrek hücre hatlarında farklı hücreler arası matriks elemanları ile kaplanmış kitosan yüzeylerde oluşturdukları sferoid yapıları mikroskop altında incelenmiştir. Ayrıca HEK293 hücrelerinin ortama salgıladıkları kollajen miktarları ELISA yöntemi ile tespit edilmiştir.Bulgular: Farklı hücreler arası matriks elemanları ve onların konsantrasyonlarının HEK293 hücrelerinin sferoid oluşumlarını farklı etkilediği tespit edilmiştir. HEK293 hücreleri yaygın olmayan ama büyük sferoid odakları oluştururken, ortamda sadece kollajen tip 1 ve jelatinin varlığında daha yaygın ve bütün yüzeye yayılım gösteren sferoidler oluşturmaktadır. Vitronektin ise sferoid oluşumunu negatif yönde etkilemektedir. Ayrıca hücreler arası matriks elemanlarının farklı dozları ve kombinasyonları ortama salınan kollajen tip 1 ve kollajen tip 4 miktarını etkilediği saptanmıştır.Sonuç: Değişen hücreler arası matriks elemanları ve konsantrasyonu 3 Boyutlu 3B kültür oluşumunu etkilemektedir. Hücrelerin sferoid oluşumunun artışını sağlayan koşulların, hücrelerden ortama salgılanan kollajen miktarını değiştirdiği de gösterilmiştir

Anahtar Kelimeler

• 3 Boyutlu Kültür, Kitosan, Kollajen, Vitronektin, Jelatin

Ertosun MG. Kitosan kaplı yüzeylerde kollajen, vitronektin ve jelatinin üç boyutlu HEK293 hücre kültürüne

Abstract

Objective: Recently, it has become very important to create a three-dimensional culture as it is easier to adapt to in vivo and clinical studies. In terms of three-dimensional culture studies, chitosan material comes to the fore because of its cost and its use in different cells. The effects of chitosan surfaces coated with extracellular matrix elements ECM on cellular spheroid formation and cellular ECM release were investigated in this study.Material and Methods: Spheroid structures formed by human embryonic kidney cell lines HEK293 on different ECM-coated chitosan surfaces were examined under a microscope. In addition, the amount of collagen released by HEK293 cells was determined by the ELISA method.Results: It was determined that the different extracellular matrix elements and their concentrations affect the spheroid formation of HEK293 cells on chitosan surfaces. While HEK293 cells form large spheroid structures with fewer foci on chitosan surfaces, they create larger spheroids in the presence of only collagen type 1 or gelatin on chitosan surfaces. Vitronectin negatively affects spheroid formation. In addition, different doses and combinations of extracellular matrix elements have been found to affect the amount of collagen type-1 and collagen type-4 released into the micro-environment.Conclusion: Changing extracellular matrix elements and their concentrations used with a chitosan surface affects 3D culture formation. It has also been shown that the conditions that increase the spheroid formation of cells change the amount of collagen secreted from cells to the micro-environment

Keywords

• 3D Culture, Chitosan, Collagen, Vitronectin, Gelatin

Kaynakça

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