This hypothesis is supported by our finding that proliferation of STAT5−/− bone marrow cells plateaued at approximately 500 pg/ml G-CSF, which is the concentration measured in the serum of STAT5−/− mice at steady state (Figure 3B).
The proliferating effect of AMD3100 on marrow cells was minimal - approximately 100-1000 folessess than that induced by G-CSF (Fig. 6B).
Sorted bone marrow cells were diluted to approximately 1.2x10 to 1.4x10cells/ml in Myelocult M5300 medium and 200 ml virus was added to the cells.
Approximately 2×10 marrow cells were plated on a 100 mm diameter Petri dish in DMEM (Dulbecco's modified Eagle's medium)/F12 (Hyclone), supplemented with 10% FBS (PAA Laboratories), 2 mM L-glutamine, 100 units/ml penicillin and 100 mg/ml streptomycin.
For example, in the TET-GFP mice after 12 weeks dox treatment, approximately 3 out of every 200 bone marrow cells were GFP+, but from the same animal, approximately 170 out of every 200 HSCs were GFP+.
Approximately 1×106 bone marrow cells were intravenously injected into C57Bl/6j mice that were previously injected with either PBS or EL4.
Approximately 2×106 bone marrow cells were cultured in a 10 cm Petri dish with 10 ml of RPMI 1640 medium plus 10% FBS, 2 mM L-glutamine, 1X Pen/Strep (complete RPMI medium) and 20% L929 cell conditioned medium.
On the day following irradiation, each animal received approximately four million bone marrow cells (BMC) that had been harvested by flushing the marrow cavity of femurs and tibiae of donor WT or C3KO mice with DMEM (Invitrogen, Carlsbad, CA) supplemented with 1% fetal calf serum (FCS; Hyclone, Logan, UT).
Approximately 5000 bone marrow cells, comprising both polychromatic erythrocytes (PCE) and normochromatic erythrocytes (NCE) were scored and the ratios between PCE and NCE were calculated.
Bone marrow cells were isolated from femur bones of approximately 8 10-week-old mice by cervical dislocation after mild anaesthesia exposure.
