Kuyu Plakalarında Dört Kanser Hücre Hattının Sferoid Oluşumu ve İnvazivlik Özelliklerinin Karşılaştırılması

Öz

Üç boyutlu (3D) hücre kültürleri, özellikle sferoidler, in vivo tümör ortamlarını taklit etmek için büyük önem taşır ve kanser araştırmaları ile ilaç testleri için daha doğru bir model sunar. Sferoidler, belirli koşullar altında hücrelerin kendi kendine bir araya gelmesiyle oluşur ve bu sayede hücresel davranışlar, istilacılık da dahil olmak üzere incelenebilir. Bu çalışma, İnsan Embriyonik Böbrek 293 (HEK-293), İnsan Kolorektal Adenokarsinom (HT-29), İnsan Meme Kanseri (MDA-MB-231) ve İnsan Serviks Kanseri (HeLa) olmak üzere dört farklı hücre hattının sferoid oluşturma yeteneğini ticari bir yuvarlak tabanlı 96 kuyucuklu mikroplak ve AggreWell™ mikrokuyucuklar kullanarak değerlendirmektedir. Hücreler, yüksek viskoziteli ortamda kültürlenmiş ve kuyucuk başına farklı hücre yoğunluklarında (1.000, 2.000 ve 3.000 hücre) ekilerek hücre sayısının zaman içinde sferoid boyutu üzerindeki etkisi incelenmiştir. Mikroskopik analizler, hücre hatları arasında belirgin farklılıklar olduğunu ortaya koymuştur; HEK-293 ve HT-29 hücreleri, daha yüksek ekim yoğunluklarında daha büyük olmak üzere, kompakt ve iyi tanımlanmış sferoidler oluşturmuştur. Buna karşılık, daha agresif ve istilacı özelliklere sahip olan MDA-231 ve HeLa hücreleri bu koşullar altında sferoid oluşturamamıştır. Bu bulgular, kanser hücrelerinin agresifliği, ekim yoğunluğu ve sferoid oluşturma yeteneği arasındaki karmaşık ilişkiyi ortaya koymakta olup, 3D kültür temelli ilaç geliştirme ve kanser araştırmalarının optimizasyonu için kritik faktörlerdir.

Anahtar Kelimeler

• 3D hücre kültürü
• Sferoid oluşumu
• Kanser hücre hatları
• Tümör invazivliği
• Hücre ekim yoğunluğu
• İlaç testi modelleri

Correlation between Spheroid Formation Ability and Reported Invasiveness of HEK-293, HT-29, MDA-MB-231, and HeLa Cancer Cell Lines in Commercial Well Plates

Öz

Three-dimensional (3D) cell cultures, such as spheroids, are essential for replicating in vivo tumor environments, offering a more accurate model for cancer research and drug testing. Spheroids form through the self-aggregation of cells under specific conditions, enabling the study of cellular behavior, including invasiveness. In this study, we investigated the correlation between spheroid formation ability and the reported invasiveness of four widely used human cancer cell lines — Human Embryonic Kidney 293 (HEK-293), Human Colorectal Adenocarcinoma (HT-29), Human Breast Cancer (MDA-MB-231), and Human Cervical Cancer (HeLa) — using a commercial round bottom 96-well microplate and AggreWell™ microwells with varying cell seeding concentrations. Cells were cultured in highly viscous media and seeded at varying densities (1,000, 2,000, and 3,000 cells per well) to assess the effect of cell number on spheroid size over time. Microscopic analysis revealed distinct differences among the cell lines; HEK-293 and HT-29 cells formed compact, well-defined spheroids, with larger spheroids observed at higher seeding densities. In contrast, the more aggressive and invasive MDA-MB-231 and HeLa cells failed to form spheroids under these conditions. These findings demonstrate the intricate relationship between cancer cell aggressiveness, seeding density, and spheroid formation ability, which are critical factors in optimizing 3D culture-based drug development and cancer research

Anahtar Kelimeler

• 3D cell culture
• Spheroid formation
• Cancer cell lines
• Tumor invasiveness
• Seeding density
• Drug testing models

Kaynakça

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