Figure 1.
Spheroid Cells Derived from Human Endometrial Cancer with High ALDH Activity Shows CSC Characteristics and Genetic Background Similar to that of the Primary Tumor
(A) H&E staining of the primary tumor (left), bright-phase image of the indicated spheroids (center left), H&E staining of xenograft tumors (center right), bright-phase image of cells grown under differentiation conditions (right). Scale bars, 100 μm.
(B) Targeted sequencing analyses of the primary tumor, spheroid cells, and spheroid-derived xenograft tumors.
(C) Western blot analyses of the spheroid cells and differentiated cells shown in (A).
(D) FACS analyses of ALDH activity after ALDEFLUOR staining. Left gated population, ALDH-low cells; right gated population, ALDH-high cells.
(E) Bright-phase images of spheroid formation (7 days after in vitro cultivation). Scale bars, 100 μm.
(F) Time course analyses of cell growth in ALDH-high and ALDH-low cells after sorting. n = 4 independent experiments, p < 0.001, Student's t tests.
(G) Western blot analyses of ALDH-high and ALDH-low cells after sorting.
(H) Gene set enrichment analyses of gene expression profiles between ALDH-high and ALDH-low cells.
(I) Time course of ALDH activity in vitro after ALDEFLUOR sorting.
(J) Volume (mean ± SEM) of xenograft tumors from 1 × 105 ALDH-low and ALDH-high cells. n = 5 independent experiments, p = 0.002, Student's t tests.
(K) Image of mice on day 33. ALDH-low cells were injected on the left side, and ALDH-high cells were injected on the right side.
(L) H&E staining and immunostaining of xenograft tumors derived from ALDH-high and ALDH-low cells, 33 days after transplantation. Scale bars, 100 μm.
(M) Limiting dilution analysis of ALDH-high cells and ALDH-low cells in vivo on day 23 after subcutaneous injection. p < 0.01.
(N) ALDH activity after ALDEFLUOR staining in cancer cells from ALDH-high and ALDH-low xenograft tumors. Experiments in (D to G) and (I to N) were performed with EMN24 cells.
∗∗ p < 0.01; ∗∗∗p < 0.001. See also Figure S1.