Intraaortic hematopoietic clusters contain pre-HSCs
| . | . | Cell number . | Neonate recipients§ . | |
|---|---|---|---|---|
| Embryo stage . | Sorted cells . | injected/recipient . | Rag2−/−γc−/− . | WT . |
| Early E10 (25-32 sp) | c-kit+* | 10 | 0/1 | 0/9 |
| 50 | — | 0/4 | ||
| 100 | 2/2 | 0/2 | ||
| 150 | — | — | ||
| 200 | — | 3‖/5 | ||
| 750 | 1¶/1 | — | ||
| 1350 | 1/1 | — | ||
| c-kit+GFP–CD45–†‡ | 1000-2500 | 3/3 | — | |
| c-kit+GFP+CD45– | 250-550 | 3/4 | — | |
| c-kit+GFP–CD45+ | 150-350 | 1/4 | — | |
| c-kit+GFP+CD45+ | 250-650 | 3/4 | — | |
| Mid-E10 (33-38 sp) | c-kit+* | 10 | 0/1 | 0/6 |
| 50 | 2/2 | 2¶#/12 | ||
| 100 | — | 1/3 | ||
| 150 | 2/2 | — | ||
| 200 | — | 1/1 | ||
| . | . | Cell number . | Neonate recipients§ . | |
|---|---|---|---|---|
| Embryo stage . | Sorted cells . | injected/recipient . | Rag2−/−γc−/− . | WT . |
| Early E10 (25-32 sp) | c-kit+* | 10 | 0/1 | 0/9 |
| 50 | — | 0/4 | ||
| 100 | 2/2 | 0/2 | ||
| 150 | — | — | ||
| 200 | — | 3‖/5 | ||
| 750 | 1¶/1 | — | ||
| 1350 | 1/1 | — | ||
| c-kit+GFP–CD45–†‡ | 1000-2500 | 3/3 | — | |
| c-kit+GFP+CD45– | 250-550 | 3/4 | — | |
| c-kit+GFP–CD45+ | 150-350 | 1/4 | — | |
| c-kit+GFP+CD45+ | 250-650 | 3/4 | — | |
| Mid-E10 (33-38 sp) | c-kit+* | 10 | 0/1 | 0/6 |
| 50 | 2/2 | 2¶#/12 | ||
| 100 | — | 1/3 | ||
| 150 | 2/2 | — | ||
| 200 | — | 1/1 | ||
—, Not done.
n = 5 independent experiments.
n = 3 independent experiments.
The doses of c-Kit+GFP–CD45–, c-kit+GFP+CD45–, c-kit+GFP–CD45+, and c-kit+GFP+CD45+ cells injected per recipient correspond to 5 to 12, 5 to 11, 4 to 9, and 5 to 13 ee injected per recipient, respectively.
Number of repopulated recipients per injected neonate recipients 4 to 5 months postinjection. Mice were considered repopulated when donor-derived cells (>0.01%) were detected in both the bone marrow and spleen by flow cytometry of the H2kk and Ly6a-GFP donor markers (for Rag2−/−γc−/− recipients) or of the CD45.1 donor marker (for WT recipients).
‖Multilineage reconstitution is shown in supplemental Figure 5.
Primary recipients used for secondary transplantations (see supplemental Table 1).
Multilineage reconstitution is shown in Figure 1H.