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First published online 24 September 2003
doi: 10.1242/dev.00755

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Mouse placenta is a major hematopoietic organ

Marcio Alvarez-Silva1,2, Patricia Belo-Diabangouaya1, Josselyne Salaün1 and Françoise Dieterlen-Lièvre1,*

1 Laboratoire d'Embryologie cellulaire et moléculaire du CNRS et du Collège de France, 49 bis, avenue de la Belle Gabrielle, 94736 Nogent s/Marne cedex, France
2 Laboratório de Neurobiologia e Hematologia Celular e Molecular, BEG, CCB, Universidade Federal de Santa Catarina, Campus Universitário, 88040-900, Florianópolis, SC, Brazil



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  		Fig. 1. E15 Placenta. Bright field, top; UV fluorescence, bottom. The fetal component is GFP+, the maternal component GFP–. Notice the zone of invasion by fetal blood vessels (weakly fluorescent between the white lines). Arrows point to the umbilical cord, the arrowhead to the zone of attachment to the uterus.

 


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  		Fig. 2. A 14-day colony from an E15 placenta from the cross between a female C57BL/6 Ly-5.2 and a C57BL/6 Ly-5.2 GFP+ male. (A) Phase contrast. (B) UV fluorescence.

 


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  		Fig. 3. Developmental frequencies of CFU-GEMMs, BFU-Es and CFU-GMs in E10 to E17 placenta, yolk sac and liver, 14 days after plating. Each point is the mean±s.d. from four independent experiments in which cells from four to six embryos were pooled (four wells in each experiment).

 


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  		Fig. 4. CFU-GEMM colonies obtained from E15 placenta, yolk sac and liver cells after 14 days of culture in methylcellulose-based medium. (A-C) Phase contrast. (D-F) May-Grünwald-Giemsa staining. (A,D) Placenta. (B,E) Yolk sac. (C,F) Liver. Placenta colonies are characterized by a very large, highly hemoglobinized center. Cytology reveals a different degree of erythroid maturity depending on the organs: early erythroblasts predominate in the placenta; late erythroblasts (small, deeply basophilic, arrows in E) are numerous in the yolk sac; fetal liver displays mainly erythrocytes expelling their nucleus (arrows in F).

 


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  		Fig. 5. Long-term HPP-CFC assay. HPP-CFCs were taken from MethoCult M3434 cultures every 20 days. 104 cells from pooled colonies were replated at each passage. HPP-CFCs were scored after a total 60 days in culture. Results are expressed as the mean of three independent observations performed in quadruplicate. This graph was established using E15 organs; similar results were yielded by E12 and E17 organs. FL, fetal liver; Pl, placenta; YS, yolk sac.

 


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  		Fig. 6. Expression of transcription factors Runx1, Scl and Tel/Etv6 in E8-E9 embryos. RT-PCR was performed on total RNA extracted from different tissues as indicated. BM, bone marrow; CH, embryo caudal halves.

 




© The Company of Biologists Ltd 2003