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Fig. 3. Replating of embryonic cells into methylcellulose. (A) E7.5
cells per embryo were co-cultured with OP-9 cells in the presence of IL3, TPO,
SCF, EPO and VEGF. On day 0, 1, 2 or 4, after treatment with trypsin, single
cells were replated into methylcellulose with IL3, TPO, SCF, EPO and VEGF.
Following 7 days of further culture, colonies formed were picked up. Cytospin
preparations were made and were stained with Hemacolor. This experiment used a
total of 20 E7.5 embryos (three MS-, four LS-, seven OB-, three EB-, three
LB-stage embryos). (B) Data from a total of four embryos are shown. Two
embryos were of ES stage and two embryos were of MS stage. nmE,
neutrophil/macrophage/erythrocyte; mE, macrophage/erythrocyte; m, macrophage.
(C) RT-PCR analysis for ßH1 and ß-major globin expression in
individual hematopoietic colonies that were formed after contact with OP-9
cells for 3 days.
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