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Turkish Journal of Biochemistry

Türk Biyokimya Dergisi

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Effect of different culture media on isolation and differentiation of dendritic cells / Farklı besiyerlerinin dendritik hücre izolasyon ve farklılaşmasına etkisi

Ayşe Mine Yılmaz
  • Department of Biochemistry, School of Medicine & Marmara University, Genetic and Metabolic Diseases Research Center, İstanbul, Turkey
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/ Ergül Mutlu Altundağ
  • Department of Biochemistry, School of Medicine & Marmara University, Genetic and Metabolic Diseases Research Center, İstanbul, Turkey
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/ Gülşah Gedik
  • Department of Biochemistry, School of Medicine & Marmara University, Genetic and Metabolic Diseases Research Center, İstanbul, Turkey
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/ Semra Koçtürk
  • Corresponding author
  • Dokuz Eylül University Faculty of Medicine, Department of Biochemistry, İzmir, Turkey & Marmara University, Genetic and Metabolic Diseases Research Center, İstanbul, Turkey
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/ A. Süha Yalçın
  • Department of Biochemistry, School of Medicine & Marmara University, Genetic and Metabolic Diseases Research Center, İstanbul, Turkey
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/ Yavuz Taga
  • Department of Biochemistry, School of Medicine & Marmara University, Genetic and Metabolic Diseases Research Center, İstanbul, Turkey
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Published Online: 2015-11-13 | DOI: https://doi.org/10.1515/tjb-2015-0031

Abstract

Objective: Dendritic cells (DCs) are members of the mammalian immune system and are considered to be the most powerful antigen presenting cells. They are responsible for the induction of T-cells or T-cell dependent immunity and tolerance. In this study we have investigated the effect of different serum supplements on generation and yield of mature dendritic cells isolated from peripheral blood mononuclear cells.

Methods: Three different serum supplements (10% Fetal Bovine Serum, 1% Human Serum Albumin and 1% autologous serum) were compared with serum-free media to identify the role and importance of serum supplements on DC cultivation. Effect of different media on maturation signs (CD40, CD80, CD86, CD209a) and cytokine release (TNF-α, IL-10, IL-12, IL-6) was examined.

Results: DCs generated in serum-free media was similar to those of cells in medium with autologous serum. Few dendritic-like cells were observed in fetal bovine serum and human serum albumin. The effect of different media on maturation of DCs was compared phenotypically and increased expression of CD80, CD86 and CD209a identified maturation and yield of DCs.

Conclusion: Our results suggest that serum free media can be used to overcome potential drawbacks associated with different serum containing supplements.

Özet

Amac: Dendritik hucreler (DC), memeli immun sistemindeki en guclu antijen sunan hucreler olarak kabul edilir. Bu hucreler, T-hucrelerin uyarılması ile T-hucre bağımlı bağışıklıktan sorumludur. Bu calışmada farklı serumların periferik kandan izole edilen mononukler hucrelerin olgun dentritik hucrelere farklılaşması uzerine etkileri araştırılmıştır.

Metod: DC kulturlerindeki serum desteklerinin rolunu tanımlamak uzere, Fetal Sığır Serumu, İnsan Serum Albumini ve Otolog serum iceren besiyerlerinin etkisi serumsuz ortam ile karşılaştırılmıştır. Olgunlaşma işaretleri (CD40, CD80, CD86, CD209a) ve sitokin salınımları (TNF-α, IL-10, IL-12, IL-6) acısından incelenmiştir.

Bulgular: Serumsuz ortamda farklılaştırılan hucreler ile otolog serum iceren hucrelerin benzer ozellikte olduğu buna karşılık fetal sığır serumu ile insan serum albumini iceren besiyerlerinin az sayıda dendritik hucre oluşturdukları gozlenmiştir. Farklı ortamların dendritik hucrelerdeki yuzey belirteclerinin etkisine bakıldığında CD80, CD86 ve CD209a artışı tanımlandı.

Sonuc: DC uretiminde farklı faktorlerin hesaba katılması gerektiği ve serumsuz ortamların daha başarılı sonuclar verdiği duşunulmektedir.

Keywords : Dendritic cells; peripheral blood mononuclear cells; serum-free media; CD209a; IL-10; IL-12

Anahtar Kelimeler : Dendritik hucreler; periferal kan mononukler hucreler; serumsuz besiyeri; CD209a; IL-10; IL-12

About the article

Received: 2015-06-02

Accepted: 2015-07-14

Published Online: 2015-11-13

Published in Print: 2015-11-01


Citation Information: Turkish Journal of Biochemistry, ISSN (Online) 1303-829X, ISSN (Print) 0250-4685, DOI: https://doi.org/10.1515/tjb-2015-0031.

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