Analysis of HSCs in BM of Lyl-1-/- by in vitro LTC-IC assay
No. plated LSK BM cells . | LTC-ICs of BM, % of negative wells . | . | No. CFCs/LTC-ICs . | . | ||
|---|---|---|---|---|---|---|
| . | Lyl-1+/+ . | Lyl-1-/- . | Lyl-1+/+ . | Lyl-1-/- . | ||
| 100 | 75 | 100 | 0 | 0 | ||
| 500 | 58 | 100 | 2.25 ± 1.2 | 0 | ||
| 1000 | 17 | 50 | 18 ± 3.5 | 3.75 ± 2 | ||
| 2500 | 0.1 | 25 | 74 ± 12 | 40 ± 8.5 | ||
No. plated LSK BM cells . | LTC-ICs of BM, % of negative wells . | . | No. CFCs/LTC-ICs . | . | ||
|---|---|---|---|---|---|---|
| . | Lyl-1+/+ . | Lyl-1-/- . | Lyl-1+/+ . | Lyl-1-/- . | ||
| 100 | 75 | 100 | 0 | 0 | ||
| 500 | 58 | 100 | 2.25 ± 1.2 | 0 | ||
| 1000 | 17 | 50 | 18 ± 3.5 | 3.75 ± 2 | ||
| 2500 | 0.1 | 25 | 74 ± 12 | 40 ± 8.5 | ||
LSK BM cells (100, 500, 1000, 2500) were plated on MS5 stroma cells and were cultured for 5 weeks; half the medium was replaced each week. At the end of the incubation time, the frequency of LTC-ICs was calculated according to Poisson law. Cells from each positive well were then plated in methylcellulose, and the number of CFC/LTC-ICs was counted after 7 days. Results are representative of 3 independent experiments (± SEM).