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Fig. 8. Effects of EPO and various growth factors on primitive and definitive
hematopoiesis. Primate ES cells were cultured for 6 days with VEGF, bFGF or
BMP4. The induced cells were harvested and re-cultured with (black circles) or
without (white circles) 10 U/ml EPO. The floating cells were analyzed as
described in Fig. 2. Adherent
clusters (
20 cells) were counted on day 10. Primitive erythroid colonies
(50 cells) were counted 7 days after replacing the medium with
methylcellulose-containing medium on day 8. (A,C) Effects of EPO and various
concentrations of VEGF (0 ng/ml, circles; 5 ng/ml, triangles; 10 ng/ml,
squares; 20 ng/ml, diamonds) on the number of erythrocytes (A) and total
hematopoietic cells (C). (B) Sequential analysis of the proportion of
definitive erythrocytes (EryD) among total erythrocytes in the presence of EPO
alone (black columns), or EPO plus VEGF (20 ng/ml; striped columns). (D)
Sequential analysis of the percentages of erythrocytes (white columns),
myeloid cells (striped columns) and megakaryocytes (black columns) among total
hematopoietic cells in the presence of EPO plus VEGF (20 ng/ml). (E,F) Effects
of EPO and growth factors on the number of adherent hematopoietic clusters (E)
and primitive erythroid colonies (F). E, EPO; V, VEGF (5 to 20 ng/ml); b, bFGF
(20 ng/ml); B, BMP4 (20 ng/ml). (G,H) Effects of bFGF (20 ng/ml, triangles),
BMP4 (20 ng/ml, squares) and VEGF (20 ng/ml, diamonds) on the number of
erythrocytes (G) and total hematopoietic cells (H). Data represent the
mean±s.d. of triplicate wells. Representative results from one of three
independent experiments are shown.
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