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. 2020 Dec 23;8:585987. doi: 10.3389/fcell.2020.585987

FIGURE 1.

FIGURE 1

Establishment of a 3D-sphere-forming culture system as an in vitro culture model of colorectal CSCs. Colorectal CSC spheres were formed using the HCT116 cancer cell line after 1 week sphere culture. The sizes of spheres greater than 100 μm were enumerated, with a representative image of a tumorsphere shown (A). Real-time PCR results demonstrating changes in the expression of the stem cell markers KLFG, NANOG, and SOX2 after 1 week in sphere culture relative to that in subconfluent monolayers (B). The results of FACS analysis showing the percentage of the total cell population that consisted of CD44+/CD133+ cells in both monolayer and sphere cultures (C). HCT116 cells were sorted by dual-color flow cytometry analysis according to CD44 and CD133 expression. The dot plot is divided into two quadrants for CD44+/CD133+ or CD44–/CD133–. The sorted HCT116 cell populations were plated into sphere-forming culture dishes, and their clonogenic abilities were analyzed (D). Real-time PCR results demonstrating changes in the expression of KLFG, NANOG, and SOX2 in both CD44+/CD133+ and CD44–/CD133– subpopulations (E). The results represent the means ± SD of three independent experiments.