Figure 4.

Explant Culture Maintains HSCs with Long-Term Multilineage Repopulating Capacity In Vivo

(A) Kinetics of human hematopoietic repopulation in the peripheral blood for 14 NSG mice transplanted with cultured human AGM (solid lines) or embryonic liver (EL) (dotted line).

(B) Representative flow cytometry plots of human myeloid (CD45⁺CD33⁺), T (CD45⁺CD3⁺), B (CD45⁺CD19⁺), NK (CD45⁺CD94⁺) cells in the peripheral blood, bone marrow, and spleen of an NSG mouse transplanted with 0.3 ee of a cultured CS16 human AGM region (part 2), culled for analysis at 7 months.

(C) Representative flow cytometry plots of HSCs (CD34⁺CD38⁻CD90⁺CD45RA⁻), multipotent progenitors (MPPs) (CD34⁺CD38⁻CD90⁻CD45RA⁻), and multi-lymphoid progenitors (MLPs) (CD34⁺CD38⁻CD90⁻CD45RA⁺) (i), erythroid cells (ii), myeloid cells (iii), NK/NKT cells (iv), and T cells (v) in the bone marrow, blood, spleen, and thymus of NSG mice. (i), (ii), and (v) are from a primary recipient mouse injected with 0.96 ee of AGM part 3 from a CS16 embryo cultured for 7 days, culled for analysis at 4 months. (iii) and (iv) are taken from a primary recipient mouse injected with 0.28 ee AGM posterior half, from a CS17 embryo cultured for 7 days, culled for analysis at 5 months.

All plots are gated on 7AAD singlets. Flow cytometry controls are shown in Figure S4. BM, bone marrow.