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Fig. 3. Assays for fetal hematopoietic stem cell (HSC) development.
(A) (Top panel) An E10.5-11 mouse embryo. HSCs can be reliably assayed
only by transplanting them into myeloablated (e.g. irradiated) recipients
(lower panel) and then documenting their ability to give rise to sustained
(3-6 months) multilineage hematopoietic reconstitution. Functional adult-type
HSCs that can achieve such reconstitution in this assay are found only in
mouse embryos or extra-embryonic tissues after E10.5-11.5. (B) (Top
panel) A mouse embryo at E9.0. HSC activity can be observed at a younger age,
and higher hematopoietic chimerism from immature HSCs (pre-HSCs) can be
obtained if more permissive recipients (e.g. newborn mice,
Rag2-/-
c-/- mice, lower left)
are used. Transplantation directly into the long bones of a recipient can also
improve engraftment (lower right). Intermediate explant culture of pSP cells
also allows detection of HSC activity at a younger age.