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Fig. 5. Myeloid and definitive erythroid potential of allantois and chorion explants. (A) FACS analysis of ten pooled allantois and nine pooled chorion explants following culture for 14 days on OP9 stromal cells in the presence of SCF, IL3, IL6, Flt3-L, G-CSF, GM-CSF and 2-ME. n_experiments=4, n_allantois=32, n_chorion=25. (B) May-Grunwald Giemsa stained nonadherent cells isolated from allantois and chorion explants cultured on OP9 cells under the conditions described in A. a-h, allantois explants; i-p, chorion explant cultures; a-c, i-n, monocytes; d-e, macrophages; f-h, o-p, myelocytes. (C) Methylcellulose colony forming assays from the allantois and chorion. Six conceptus equivalents of allantoises and chorions were cultured together as explants, disaggregated and plated in methycellulose. The top three panels are representative colonies. (Bottom) May-Grunwald Giemsa-stained cytospin preparations of individual colonies from the allantois and chorion cultures. e, erythryocyte; m, myeloid cell; bar=10 µm. Colony numbers per six conceptus equivalents (±s.e.m. in parentheses) following 2 days of explant culture in the presence of 50% rat serum on plastic, or after 5 days of explant culture on OP9 cells in the presence of SCF, Flt3-L and EPO were averaged from three independent experiments. (D) RT-PCR for globin gene expression (y and ßmajor). BFU-E or CFU-GM represents individual colonies from either the allantois or chorion methylcellulose cultures. Yolk sac was isolated from 9.0 dpc conceptuses as a positive control for y expression, and bone marrow was used as a negative control.

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