Ayşe EROL, Figen ABATAY SEL, Mediha SÜLEYMANOĞLU, Gökhan DEMİRAYAK, Dürdane Serap KURUCA, Fatma Savran OĞUZ

KORDON KANI VE WHARTON JELİ KAYNAKLI MEZENKİMAL KÖK HÜCRELERİN İMMUNOMODÜLATÖR ÖZELLİKLERİNİN KARŞILAŞTIRILMASI

Amaç: Mezenkimal kök hücreler (MKH’ler), immün düzenleyici özellikleri nedeniyle bir çok immün sistem ilişkili hastalık için tedavi seçeneği olarak umut vaad etmektedir. Wharton Jeli (WJ-MKH) ve kordon kanı (KK-MKH), diğer MKH kaynaklarına göre son dönemlerde daha çok ilgi görmektedir. Çalışmada, WJ-MKH’ler ve KK-MKH’lerin immünolojik etkilerinin mekanizmalarındaki farkı araştırmak amaçlanmıştır. Gereç ve Yöntem: Periferik kan mononükler hücreler (PKMH) ve MKH ko-kültür öncesi ve sonrası CD4+ T hücrelerin hücre içi sitokin seviyeleri (Interlökin-4 (IL-4), Interferon-γ (IFN-γ), and Interlökin-17 (IL-17)) flow sitometri ile tespit edildi. Aynı zamanda Tümör büyüme faktörü (TGF)-β, IL-4, IL-17, IFN-γ süpernatant sitokin seviyeleri ELISA ile ölçüldü. Çalışmada, ko-kültür MKH/PKMH 1/5 ve 1/10 oranlarında, 24 saat ve 72 saat inkübasyon süreleri uygulandı. Bulgular: Verilerimiz WJ-MKH’lerin ve KK-MKH’lerin farklı morfolojik özellikleri, çoğalma kapasiteleri, çoğalma süreleri ve immünomodülasyon yetenekleri olduğunu gösterdi. IFN-γ, KK-MKH/PKMH ko-kültür sonrası WJ-MKH/PKMH ko-kültürü sonrasına göre hücre içerisinde hem 1/5 hem 1/10 oranlarında 24. saatlerde anlamlı derecede azalırken, 72. saatlerde anlamlı derecede arttı (p<0,05). IL-17 sitokinin için farklı olarak sadece 72. saatte KK-MKH grubunda WJ-MKH grubuna göre anlamlı derecede hücre içi seviyesi düştü. IL-4 için KK-MKH grubunda 1/10 oran 24 saatte hücre içi seviyesi anlamlı derecede düşerken diğer tüm gruplarda artış gösterdi. KK-MKH/PKMH ko-kültür sonrası TGF-β supernatant seviyesi 1/5 oranında 24 saatte azalırken 72 saatte artış gösterdi. 1/10 oranlarında ise tam tersi 24. saatte KK-MKH grubunda artarken 72 saatlerde anlamlı derecede azaldı (p<0,05). Sonuç: KK-MKH‘ler ve WJ-MKH’ler, farklı immünmodülatör özellikler göstermektedir. Bu bulgular, KK-MKH’lerin izolasyonun, çoğalma kapasitesinin ve immünmodülatör özelliklerinin farklı olması sebebiyle hücre tedavileri açısından WJ-MKH’lerin kullanılmasının daha avantajlı olduğu söylenebilir.

Anahtar Kelimeler:

Kordon kanı, Kordon dokusu, mezenkimal kök hücre, PKMH, ko-kültür, immunmodülasyon

COMPARISON OF THE IMMUNOMODULATORY PROPERTIES OF THE CORD BLOOD AND WHARTON’S JELLY DERIVED MESENCHYMAL STEM CELLS

Objective: Mesenchymal stem cells (MSCs) are promising as a treatment option for many immune-related diseases due to their immune regulatory properties. Wharton’s Jelly (WJ-MSC) and cord blood (CB-MSC) have recently received more attention than the other MSC sources. In this study, it was aimed to investigate the difference in the mechanisms of the immunological effects of WJ-MSCs and CB-MSCs. Material and Methods: The intracellular cytokine levels of peripheral blood mononuclear cells (PBMC) and CD4+ T cells before and after MSC co-culture (Interleukin-4 (IL-4), Interferon-γ (IFN-γ), and Interleukin-17 (IL17)) were determined by flow cytometry. At the same time, tumor growth factor (TGF)-β, IL-4, IL-17, IFN-γ supernatant cytokine levels were measured by ELISA. In the study, incubation times of 24 hours and 72 hours were applied with co-culture MSC/PBMC ratios of 1/5 and 1/10. Results: Our data showed that WJ-MSCs and CB-MSCs have different morphological features, proliferation capacities, proliferation times and immunomodulating abilities. One of the cytokines of IFN-γ decreased significantly at both 1/5 and 1/10 ratios in the cell at 24 hours and increased significantly at 72 hours after CB-MSC/PBMC co-culture compared to the level after WJ-MSC/PBMC co-culture (p<0.05). Unlike for IL-17 cytokine, the intracellular level decreased significantly in the CB-MSC group only at 72 hours compared to the WJ-MSC group. For IL-4, the 1/10 ratio in the CB-MSC group decreased significantly at 24 hours, while the intracellular level was increased in all other groups. After CB-MSC/PBMC co-culture, TGF-β supernatant level decreased by 1/5 in 24 hours and increased in 72 hours. On the contrary, TGF-β supernatant level increased in the CB-MSC group at 24 hours and decreased significantly at 72 hours in 1/10 ratios (p<0.05). Conclusion: CB-MSCs and WJ-MSCs show different immunomodulatory properties. Based on these findings, it can be said that the use of WJ-MSCs is more advantageous in terms of cell therapies due to the different isolation, proliferation capacity and immunomodulatory properties of CB-MSCs.

Keywords:

Cord blood, Cord tissue, MSCs, Immunomodulation,

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