Figure 3.

In vitro hematopoietic support capacity of stromal cell fractions. Stromal fractions were isolated from collagenase treated spleen stroma, sorted on the basis of marker expression and cultured for 28 days until confluent. Lin^- bone marrow cells was then overlaid above stroma at 1–5 × 10⁴ cells/ml in 20 ml. Non-adherent cells were collected after 21 days and stained with antibodies to CD11b, CD11c, MHC-II and F4/80 to delineate myeloid/dendritic cell production. Flow cytometric analysis was used to identify subsets of progenitors (CD11b^-CD11c^-), myeloid cells (CD11b⁺CD11c^-F4/80^-MHC-II^-), L-DC (CD11b⁺CD11c⁺F4/80⁺MHC^-II^-), and cDC-like cells (CD11b^+/-CD11c⁺F4/80^-MHC^-II⁺). Multiple individual self-controlled experiments were conducted, each comparing cell production by different fractions in comparison with 5G3. Data is shown for each of 9 experiments as: (A) % yield as number of cells produced relative to input cell number, and (B) proportional representation of subsets.