Selinay ŞENKAL, Derya BURUKÇU, Engin SÜMER, Fikrettin ŞAHİN, Ayşegül DOĞAN, Taha Bartu HAYAL, Binnur KIRATLI, Hatice Burcu ŞİŞLİ, Derya SAĞRAÇ, Ayla Burçin ASUTAY

Ex vivo dermal toksikoloji analizleri, in vivo test yöntemlerinin yerini alabilmesi için çok önemlidir ve son yıllarda kozmetik, ilaç ve kimyasalların test edilmesi için ilgi çekici hale gelmiştir. Kültürde yapılandırılmış uygun bir epidermis modelinin geliştirilmesi, tek katmanlı kültür sistemlerinin sınırlamalarını ortadan kaldırabilir. Bu amaçla, mevcut çalışmada, keratinosit bazlı toksisite analizi amacı ile bir ex vivo 3D hücre kültürü sistemi geliştirmek için ölümsüzleştirilmiş insan keratinosit hücre hattı (HaCaT) kullandık. Fare embriyonik fibroblast kondisyonel besiyeri ve Matrigel matriksi tabanlı 3D HaCaT hücre kültürü sistemleri, kültürde deri ilgili genleri ve proteinleri ifade etti. 3D HaCaT kültürleri, deri benzeri bir fenotip gösterdi ve seçilen test bileşiklerine karşı tepki gösterdi. 1 saat ve 24 saat boyunca seçilen kimyasallara maruz bırakılan tek katmanlı HaCaT hücrelerine kıyasla güvenilir sonuçlar elde edildi. 3D HaCaT hücre kültürlerinin ve tek tabakalı kültürlerin gen ekspresyon profilleri, test bileşiklerinin uygulanmasından sonra tamamen farklıydı.Özet olarak, sonuçlarımız Matrigel matrisinde oluşturulan bir 3D HaCaT hücre kültürü sisteminin cilt epidermisi benzeri bir fenotip uyguladığını gösterdi. Sonuç olarak, 3D HaCaT hücre kültürleri, in vitro toksikoloji deneylerinin yürütülmesi için kabul edilebilir bir test yöntemi olabilir.

3D CULTURE OF HaCaT KERATINOCYTE CELL LINE AS AN in vitro TOXICITY MODEL

Ex vivo dermal toxicology analyses are crucial for replacement of in vivo test methods and have been of interest in recent years for testing cosmetics, drugs, and chemicals. Development of an appropriate reconstructed epidermis model might overcome the limitations of monolayer culture systems. In the current study, we used the immortalized human keratinocyte cell line (HaCaT) to develop an ex vivo 3D cell culture system for keratinocyte-based toxicity analysis. Mouse embryonic fibroblast-conditioned medium and Matrigel matrix-based 3D HaCaT cell culture systems expressed skin-related genes and proteins in culture. The 3D HaCaT cultures demonstrated a skin-like phenotype and response against selected test compounds. Reliable results were obtained compared to monolayer HaCaT cells which were exposed to selected chemicals for 1 h and 24 h. Gene expression profiles of 3D HaCaT cell cultures and monolayer cultures were completely different after administration of the test compounds.Overall, our results showed that a 3D HaCaT cell culture system generated in Matrigel matrix exerted a skin epidermis-like phenotype. Consequently, 3D HaCaT cell cultures may be an acceptable test method for conducting in vitro toxicology experiments.

Keywords:

skin irritation, epidermis, Reconstructed skin, 3D skin model,

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Trakya University Journal of Natural Sciences-Cover

ISSN: 2147-0294
Yayın Aralığı: Yılda 2 Sayı
Başlangıç: 2000
Yayıncı: Trakya Üniversitesi

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