Table 1. Tumor-initiating cells were enriched in Lin−CD29HCD24H subpopulation.
Cells from T6, T7, T10 (primary tumor), T1–T5, T8, T9 (transplantation generation 1, TG1), were freshly digested and FACS sorted. All dead cells were gated out with PI. Mouse lineage panel plus anit-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-week-old Balb/c female mice. Mice were monitored until tumors were observed or up to 18 months if no tumors were detected. Tumor-initiating cell frequency and the Poisson distribution analysis was 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 weeks. 1,500 of Lin−CD29HCD24H and Lin−CD29HCD24L cells formed palpable tumors from 5 to 8 weeks, similar to tumors formed from 2,500 Lin−CD29LCD24L, Lin−CD29LCD24H and Lin− cells. Palpable tumors from 500 cells took 7 to 11 weeks, while those from 100 cells took 8 to 15 weeks. 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. Table 1
30,000 | 5,000 | 2,500* | 1,500** | 500 | 100** | Tumor-initiating cell frequency (95% CI) | |
---|---|---|---|---|---|---|---|
Lin− CD29HCD24H | 36/36 | 9/10 | 44/52 | 8/14 | 1/302 (1/413 - 1/221) | ||
Lin− CD29HCD24L | 35/40 | 4/11 | 16/54 | 0/12 | 1/2156 (1/2934 - 1/1584) | ||
Lin− CD29LCD24H | 7/44 | 1/4 | 0/10 | 1/14** | 0/6 | 1/25891 (1/49897 - 1/13434) | |
Lin− CD29LCD24L | 2/44 | 1/4 | 0/10 | 0/13** | 0/6 | 1/81931 (1/254398 - 1/26387) | |
Lin− | 8/10 | 3/18 | 1/4 | 0/10 | 1/21583 (1/38743 - 1/12023) |