Figure 1.

Phenotypes of BM cells cultured under various conditions. Unfractionated BM cells (10⁵) from C57BL/6 mouse were plated onto the OP9 stroma cells and cultured in mSFO2 containing 10% FCS (A), 10 ng/ml bFGF, and 100 ng/ml SCF (B), or 30 ng/ml EGF and 100 ng/ml SCF (C) in 60-mm culture dishes for 14 d. (D) Unfractionated BM cells (10⁵) were plated onto OP9 stromal cells transformed with the v-erbB2 gene (OP9/erbB2) in the mSFO2 plus SCF (100 ng/ml) for 21 d. Cultured BM cells were analyzed by May-Gruenwald-Giemsa staining (left) and by flow cytometry (right) after staining with biotinylated anti–c-Kit (ACK4), FITC-conjugated Mac-1, and anti-B220 antibody. Biotinylated ACK4 was detected with streptavidin-PE. The numbers in each quadrant represent the percentages of gated bone marrow cells. Bar, 10 μm.