Effect of BMP4 and TPO on cell ploidization and platelet production from CD34+ cells
| . | Day 3 . | Day 7 . | Day 10 . | Day 18 . | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| None . | TPO . | BMP4 . | None . | TPO . | BMP4 . | None . | TPO . | BMP4 . | None . | TPO . | BMP4 . | ||
| Ploidy | |||||||||||||
| Sub G1 | 15 ± 11 | 1.5 ± 1 | 2 ± 2 | 8 ± 4 | 5 ± 4 | 5 ± 3 | 11 ± 4 | 5 ± 2 | 5 ± 3 | 20 ± 10 | 9 ± 7 | 10 ± 5 | |
| 2N | 74 ± 7 | 77 ± 8 | 74 ± 9 | 79 ± 4 | 69 ± 4 | 68 ± 3 | 71 ± 4 | 66 ± 4 | 65 ± 3 | 62 ± 7 | 66 ± 7 | 62 ± 6 | |
| 4N | 8.5 ± 9 | 19 ± 8 | 22 ± 8 | 11 ± 3 | 21 ± 3 | 22 ± 4 | 14 ± 3 | 22 ± 2 | 22 ± 3 | 13 ± 3 | 19 ± 2 | 19 ± 2 | |
| 8N | 2 ± 1 | 2 ± 1 | 1.5 ± 0.5 | 2 ± 0.3 | 4.5 ± 1 | 4 ± 1 | 3 ± 1 | 5 ± 1 | 6 ± 1 | 3 ± 1 | 4 ± 1 | 5 ± 1 | |
| > 16N | 0.5 ± 0.1 | 0.5 ± 0.1 | 0.5 ± 0.1 | 0.5 ± 0.1 | 0.5 ± 0.1 | 1 ± 0.1 | 1 ± 0.5 | 2 ± 0.5 | 2 ± 1 | 2 ± 1 | 2 ± 1 | 4 ± 2 | |
| n | 3 | 3 | 3 | 9 | 9 | 9 | 8 | 8 | 8 | 5 | 5 | 5 | |
| Platelet ×106/106 initial CD34+ | 10 ± 2.5 | 61 ± 25 | 31 ± 8 | 39 ± 22 | 64 ± 26 | 55 ± 24 | 53 ± 37 | 186 ± 56 | 135 ± 58 | 96 ± 36 | 294 ± 160 | 196 ± 30 | |
| . | Day 3 . | Day 7 . | Day 10 . | Day 18 . | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| None . | TPO . | BMP4 . | None . | TPO . | BMP4 . | None . | TPO . | BMP4 . | None . | TPO . | BMP4 . | ||
| Ploidy | |||||||||||||
| Sub G1 | 15 ± 11 | 1.5 ± 1 | 2 ± 2 | 8 ± 4 | 5 ± 4 | 5 ± 3 | 11 ± 4 | 5 ± 2 | 5 ± 3 | 20 ± 10 | 9 ± 7 | 10 ± 5 | |
| 2N | 74 ± 7 | 77 ± 8 | 74 ± 9 | 79 ± 4 | 69 ± 4 | 68 ± 3 | 71 ± 4 | 66 ± 4 | 65 ± 3 | 62 ± 7 | 66 ± 7 | 62 ± 6 | |
| 4N | 8.5 ± 9 | 19 ± 8 | 22 ± 8 | 11 ± 3 | 21 ± 3 | 22 ± 4 | 14 ± 3 | 22 ± 2 | 22 ± 3 | 13 ± 3 | 19 ± 2 | 19 ± 2 | |
| 8N | 2 ± 1 | 2 ± 1 | 1.5 ± 0.5 | 2 ± 0.3 | 4.5 ± 1 | 4 ± 1 | 3 ± 1 | 5 ± 1 | 6 ± 1 | 3 ± 1 | 4 ± 1 | 5 ± 1 | |
| > 16N | 0.5 ± 0.1 | 0.5 ± 0.1 | 0.5 ± 0.1 | 0.5 ± 0.1 | 0.5 ± 0.1 | 1 ± 0.1 | 1 ± 0.5 | 2 ± 0.5 | 2 ± 1 | 2 ± 1 | 2 ± 1 | 4 ± 2 | |
| n | 3 | 3 | 3 | 9 | 9 | 9 | 8 | 8 | 8 | 5 | 5 | 5 | |
| Platelet ×106/106 initial CD34+ | 10 ± 2.5 | 61 ± 25 | 31 ± 8 | 39 ± 22 | 64 ± 26 | 55 ± 24 | 53 ± 37 | 186 ± 56 | 135 ± 58 | 96 ± 36 | 294 ± 160 | 196 ± 30 | |
Isolated CD34+ cells were incubated in the 4GF serum-free medium in the presence or not of 100 ng/mL of BMP4 or TPO. After 3, 7, 10, or 18 days, cells were counted and assessed for ploidization by flow cytometry using a Facscalibur cell analyzer (BD Biosciences). Results are expressed as mean cells in each ploidy class, and the number of experiments for each time point is indicated as n experiments. The numbers of platelets are expressed as the total number of platelets ×106 per 106 initial CD34+ and represent the mean of 3 independent experiments.