Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor
Yıl: 2023 Cilt: 14 Sayı: 2 Sayfa Aralığı: 108 - 118 Metin Dili: İngilizce DOI: 10.31067/acusaglik.1225228 İndeks Tarihi: 01-06-2023
Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor
Öz: Introduction&Objective: Breast cancer is the most frequently diagnosed cancer and the leading cause of death from cancer among females worldwide. Breast cancer tumors that feature breast cancer stem cells (BCSCs) are known to cause drug resistance and metastasis. Culturing BCSCs from primary tumors as mammospheres is both difficult and costly. This study aims to present an optimized mammosphere formation assay protocol and improve the breast cancer stem cell characterization process by determining the appropriate mammosphere forming method and proper cell density of cancer stem cells isolated from breast tumors. Material&Method: Cancer stem cells were isolated from breast tumors of two patients with invasive ductal carcinoma (IDC) and culturated. Subsequently, performed specific BCSCs surface markers and ALDH analysis by flow cytometry. Two mammosphere forming methods, i.e., low attachment and agar-coated wells seeded in three different cell concentrations, were compared. Results: CD44+, CD24- and CD133+ antibody expressions showed that these cells could be tumor-initiating CSCs. ALDH assay results also indicated that these cells possessed stem cell features. In addition, the results of the mammosphere assay revealed that agar-coated wells at a concentration of 7000 cells/cm2 had more prominent floating features and viable spheres. Conclusion: The findings of this study supported the hypothesis that agar-coated culture plates in mammosphere culture would increase the mammosphere formation efficiency (MFE) value and revealed the importance of the number of cells in elucidating the nature of BCSCs.
Anahtar Kelime: Primer Meme Tümörü Kanser Kök Hücre Karakterizasyonunda Mamosfer Oluşturma Optimizasyonu
Öz: Giriş&Amaç: Meme kanseri, dünya çapında en sık teşhis edilen kanserdir ve kadınlar arasında kansere bağlı ölümlerin önde gelen nedenidir. Meme kanseri kök hücrelerini (MKKH) içeren meme tümörlerinin, ilaç direncine ve metastaza neden olduğu bilinmektedir. Primer tümörlerden MKKH’ lerin mamosfer olarak kültürlenmesi hem zor hem de maliyetlidir. Bu çalışma, meme tümörlerinden izole edilen kanser kök hücrelerinden, uygun mamosfer oluşturma tekniğini ve uygun hücre yoğunluğunu belirleyerek optimize edilmiş bir mamosfer oluşturma protokolü sunmayı ve meme kanseri kök hücre karakterizasyon sürecini iyileştirmeyi amaçlamaktadır. Gereç&Yöntem: İnvaziv duktal karsinomalı iki hastanın meme tümörlerinden kanser kök hücrelerinin izolasyonu ve kültürü gerçekleştirildi. Daha sonra, akım sitometrisi ile spesifik MKKH’leri yüzey belirteçleri saptandı ve ALDH analizi gerçekleştirildi. Düşük tutunma özellikli ve agar kaplanmış kuyucuklara üç farklı konsantrasyonda hücre ekilerek iki mamosfer oluşturma yöntemi karşılaştırıldı. Bulgular: CD44+, CD24- ve CD133+ antikor ifadeleri, bu hücrelerin tümör başlatıcı KKH’leri olabileceğini gösterdi. ALDH analizi sonuçları da bu hücrelerin kök hücre özelliklerine sahip olduğunu gösterdi. Ek olarak, mamosfer testinin sonuçları, 7000 hücre/cm2’lik bir konsantrasyonda agar kaplı kuyucukların daha belirgin yüzen özelliklere ve canlı sferlere sahip olduğunu ortaya çıkardı. Sonuç: Bu çalışmanın bulguları, mamosfer kültüründe agar kaplı kültür kaplarının mammosphere oluşum etkinliği (MFE) değerini artıracağı hipotezini desteklemiş ve MKKH’ lerin doğasını aydınlatmada hücre sayısının önemini ortaya koymuştur.
Anahtar Kelime: Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık
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| APA | Uslu G, unal halbutogullari z, GACAR G, Utkan N, Yazir Y (2023). Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. , 108 - 118. 10.31067/acusaglik.1225228 |
| Chicago | Uslu Gizem,unal halbutogullari zehra seda,GACAR GÜLÇİN,Utkan Nihat Zafer,Yazir Yusufhan Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. (2023): 108 - 118. 10.31067/acusaglik.1225228 |
| MLA | Uslu Gizem,unal halbutogullari zehra seda,GACAR GÜLÇİN,Utkan Nihat Zafer,Yazir Yusufhan Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. , 2023, ss.108 - 118. 10.31067/acusaglik.1225228 |
| AMA | Uslu G,unal halbutogullari z,GACAR G,Utkan N,Yazir Y Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. . 2023; 108 - 118. 10.31067/acusaglik.1225228 |
| Vancouver | Uslu G,unal halbutogullari z,GACAR G,Utkan N,Yazir Y Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. . 2023; 108 - 118. 10.31067/acusaglik.1225228 |
| IEEE | Uslu G,unal halbutogullari z,GACAR G,Utkan N,Yazir Y "Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor." , ss.108 - 118, 2023. 10.31067/acusaglik.1225228 |
| ISNAD | Uslu, Gizem vd. "Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor". (2023), 108-118. https://doi.org/10.31067/acusaglik.1225228 |
| APA | Uslu G, unal halbutogullari z, GACAR G, Utkan N, Yazir Y (2023). Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi, 14(2), 108 - 118. 10.31067/acusaglik.1225228 |
| Chicago | Uslu Gizem,unal halbutogullari zehra seda,GACAR GÜLÇİN,Utkan Nihat Zafer,Yazir Yusufhan Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi 14, no.2 (2023): 108 - 118. 10.31067/acusaglik.1225228 |
| MLA | Uslu Gizem,unal halbutogullari zehra seda,GACAR GÜLÇİN,Utkan Nihat Zafer,Yazir Yusufhan Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi, vol.14, no.2, 2023, ss.108 - 118. 10.31067/acusaglik.1225228 |
| AMA | Uslu G,unal halbutogullari z,GACAR G,Utkan N,Yazir Y Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi. 2023; 14(2): 108 - 118. 10.31067/acusaglik.1225228 |
| Vancouver | Uslu G,unal halbutogullari z,GACAR G,Utkan N,Yazir Y Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi. 2023; 14(2): 108 - 118. 10.31067/acusaglik.1225228 |
| IEEE | Uslu G,unal halbutogullari z,GACAR G,Utkan N,Yazir Y "Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor." Acıbadem Üniversitesi Sağlık Bilimleri Dergisi, 14, ss.108 - 118, 2023. 10.31067/acusaglik.1225228 |
| ISNAD | Uslu, Gizem vd. "Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor". Acıbadem Üniversitesi Sağlık Bilimleri Dergisi 14/2 (2023), 108-118. https://doi.org/10.31067/acusaglik.1225228 |
