American Society of Hematology

Figure 2

$Figure 2. Epo and hypoxia promote the transition from expanding stress erythroid to progenitors capable of differentiation. (A) Unfractionated bone marrow cells were plated in IMDM supplemented with BMP4 + Shh + GDF15 + SCF + Epo and cultured at 2%O2 (2% O2 + Epo or SEDM) or media lacking Epo cultured at 20% O2 (20% O2 − Epo or SEEM). After 5 days of culture, 5 × 104 cells were plated in methylcellulose media containing BMP4 + SCF + Epo at 2% O2 for 5 days. Stress BFU-Es were scored after benzidine staining. (B) Unfractionated bone marrow cells were cultured in SEEM for 5 days and then switched into SEEM supplemented with Epo and cultured at 20% O2 or with Epo and cultured at 2% O2 for the indicated days. On the indicated days, 5 × 104 cells were plated in methylcellulose media supplemented with Epo, and stress BFU-Es were scored by benzidine staining after 5 days of culture. The data represent at least 2 independent experiments.$

Epo and hypoxia promote the transition from expanding stress erythroid to progenitors capable of differentiation. (A) Unfractionated bone marrow cells were plated in IMDM supplemented with BMP4 + Shh + GDF15 + SCF + Epo and cultured at 2%O₂ (2% O₂ + Epo or SEDM) or media lacking Epo cultured at 20% O₂ (20% O₂ − Epo or SEEM). After 5 days of culture, 5 × 10⁴ cells were plated in methylcellulose media containing BMP4 + SCF + Epo at 2% O₂ for 5 days. Stress BFU-Es were scored after benzidine staining. (B) Unfractionated bone marrow cells were cultured in SEEM for 5 days and then switched into SEEM supplemented with Epo and cultured at 20% O₂ or with Epo and cultured at 2% O₂ for the indicated days. On the indicated days, 5 × 10⁴ cells were plated in methylcellulose media supplemented with Epo, and stress BFU-Es were scored by benzidine staining after 5 days of culture. The data represent at least 2 independent experiments.

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