Table 1.

Tumor-initiating cells were enriched in LinCD29HCD24H subpopulation

30,0005,0002,500 *1,500 500100 Tumor-initiating cell frequency (95% confidence interval)
LinCD29HCD24H36/369/1044/528/141/302 (1/413–1/221)
LinCD29HCD24L35/404/1116/540/121/2,156 (1/2,934–1/1,584)
LinCD29LCD24H7/441/40/101/14 0/61/25,891 (1/49,897–1/13,434)
LinCD29LCD24L2/441/40/100/13 0/61/81,931 (1/254,398–1/26,387)
Lin8/103/181/40/101/21,583 (1/38,743–1/12,023)
  • NOTE: Cells from T6, T7, T10 (primary tumor), T1-T5, T8 and T9 (transplantation generation 1, TG1) were freshly digested and FACS sorted. All dead cells were gated out with PI. Mouse lineage panel plus anti-CD31 were used to exclude mouse lineage-positive cells for tumors T1-T5, T8, and T9. Anti-CD140a was added as a lineage marker for tumors T6, T7, and T10. After FACS sorting, cells were washed with PBS and transplanted into the cleared fat pad of 3-wk-old BALB/c female mice. Mice were monitored until tumors were observed or up to 18 mo if no tumors were detected. Tumor-initiating cell frequency and the Poisson distribution analysis were generated using R software (The R Foundation for Statistical Computing).

  • * T6 (primary), T4 (TG1).

  • T6, T7, T10 (primary), T1, T2 (TG1). Subpopulation-derived palpable tumors from 5,000 cells were detected between 4 to 6 wk. 1,500 of LinCD29HCD24H and LinCD29HCD24L cells formed palpable tumors from 5 to 8 wk, similar to tumors formed from 2,500 LinCD29LCD24L, LinCD29LCD24H, and Lin cells. Palpable tumors from 500 cells took 7 to 11 wk, whereas those from 100 cells took 8 to 15 wk. All primary tumors, TG1 tumors, and subpopulation-derived tumors were excised for FACS and immunostaining analysis between diameter(s) of 1 to 1.5 cm.