Chapter title |
Three-Dimensional Cell Culture Model Utilization in Renal Carcinoma Cancer Stem Cell Research
|
---|---|
Chapter number | 6 |
Book title |
Epithelial Cell Culture
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8600-2_6 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8599-9, 978-1-4939-8600-2
|
Authors |
Kamila Maliszewska-Olejniczak, Klaudia K. Brodaczewska, Zofia F. Bielecka, Anna M. Czarnecka, Maliszewska-Olejniczak, Kamila, Brodaczewska, Klaudia K., Bielecka, Zofia F., Czarnecka, Anna M. |
Abstract |
Specific 3D conditions of cancer cell lines have been optimized over last years, with growing significance of serum-free and xeno-free culture variants. The choice of proper culture media enables cancer stem cells proliferation in primary and stable cell lines. To obtain renal cell cancer stem-like phenotype, we employed media dedicated for mesenchymal cells and adult stem cells. Developed RCC cell line 3D culture system enables effective drug testing, including tyrosine kinase inhibitor anti-cancer cell toxicity. To induce formation of 3D spheroids by RCC cell lines, StemXvivo and NutriStem media must be used. Usage of laminin- or poly-D-lysine coated plates enhances also the formation of spheroids in 3D-promoting media. Seeding is optimal with Caki-1 or ACHN cell lines as well as 786-O or HKCSC cells. Our bio-mimic 3D RCC cell culture model promotes cell viability and stem-related gene expression including E-cadherin, N-cadherin, HIF1, HIF2, VEGF, Sox2, Pax2, and Nestin. 3D spheroid formation ability and spheroid volume increase are disturbed upon drug treatment. Untreated 3D structures reach ~100 μm in diameter at the end of 14-day long experiment. Sorter-based cell cycle analysis and Ki-67 staining should be conducted to verify specific toxicity. We suggest that due to the more complex architecture 3D RCC culture is more relevant to investigate the in vivo-like tumor drug response. |
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