In Vitro Murine Hematopoiesis Supported by Signaling from a Splenic Stromal Cell Line

Figure 3

Characterization of cells produced in stromal cocultures of Lin⁻ bone marrow cells over 5G3 stroma. (a) Production of nonadherent cells at 14 and 21 days. Nonadherent cells were collected from the supernatant and stained with antibodies specific for CD11b, CD11c, MHC-II, and F4/80 for flow cytometric analysis to delineate subset type. L-DC was gated as CD11b⁺CD11c^+/−F4/80⁺MHC-II⁻, cDC-like cells as CD11b⁺CD11c^+/−F4/80⁻MHC-II⁺, and immature myeloid cells as CD11b⁺CD11c^+/−F4/80⁻MHC-II⁻. Progenitors were enriched in the CD11b⁻CD11c⁻F4/80⁻MHC-II⁻ subset. One coculture representative of multiple experiments is shown. (b) Diagrammatic representation of cell types characterized in cocultures of Lin⁻ bone marrow over 5G3 stroma.

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