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. 2019 Jan 11;54(3):893–904. doi: 10.3892/ijo.2019.4683

Figure 3.

Figure 3

Organoid culture maintains the proportion of cancer stem cells during serial passages. (A) Organoids or spheres generated from purified CD133+ and CD133− CRC cells. ***P<0.001. Scale bars, 400 µm. (B) Reverse transcription-quantitative polymerase chain reaction analysis of the expression of CD133 and CD44 at the mRNA level in organoid- and sphere-derived cells during serial passages. *P<0.05 and ***P<0.001. (C) Immunofluorescence staining of CD133 and CK20 in organoids during serial passages. Scale bars, 400 µm. (D) Immunofluorescence staining of CD133 and CK20 in spheres during serial passages. Scale bars, 400 µm. (E) Quantification analysis of CD133+ cells in organoids and spheres during serial passages. *P<0.05. (F) Flow cytometric analysis of CK20+ cells in organoids and spheres during serial passages. *P<0.05, **P<0.01 and ***P<0.001. The experiments in (A-F) were duplicated in cells derived from CRC5, CRC6 and CRC9 (n=3); the data obtained from CRC6 are presented here. CRC, colorectal cancer; CD133, cluster of differentiation 133; CK20, cytokeratin 20; NS, not significant.