Clonal Analysis of Mouse Development Reveals a Polyclonal Origin for Yolk Sac Blood Islands

Institute of Stem Cell Biology and Regenerative Medicine and Department of Pathology and Developmental Biology, Stanford University School of Medicine, Stanford, California 94305, USA.
Developmental Cell (Impact Factor: 9.71). 11/2006; 11(4):519-33. DOI: 10.1016/j.devcel.2006.08.001
Source: PubMed


Direct clonal analysis of tissue and organ maturation in vivo is a critical step in the interpretation of in vitro cell precursor-progeny relationships. We have developed a method to analyze clonal progenitor contributions in vivo using ES cells stably expressing separate fluorescent proteins and placed into normal blastocysts to form tetrachimeras. Here we applied this method to the analysis of embryonic yolk sac blood islands. In most vertebrates, yolk sac blood islands are the initial sites of appearance of hematopoietic and endothelial cells. It has been proposed that these lineages arise from a common clonal progenitor, the hemangioblast, but this hypothesis has not been tested directly in physiological development in vivo. Our analysis shows that each island has contributions from multiple progenitors. Moreover, contribution by individual hemangioblast progenitors to both endothelial and hematopoietic lineages within an island, if it happens at all, is an infrequent event.

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Available from: Hiroo Ueno,
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    • "Mice were sacrificed, and the testes were fixed in 4% PFA at 4°C overnight, frozen in OCT compound, cut, and analyzed as reported previously3435. Immunostaining was performed using the following primary antibody with anti-GFRα1 (R&D Systems, Minneapolis, MN, USA) or anti-PLZF (Santa Cruz, Dallas, TX, USA) antibody followed by Alexa Fluor 594- or 750-labeled secondary antibodies (Molecular Probes, Eugene, OR, USA). Nuclear counter-staining was performed using Hoechst 33342 (Sigma) as described previously3435. "
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    • "In these post-MET stages, tubules were polyclonal, derived from multiple mixed progenitors (Figures 3B–3D, dotted white lines) similar to lineage-tracing observations of adult kidneys. We separately generated tetrachimeric mice by injecting mouse embryonic stem cells (mESCs) that stably express separate fluorescent proteins (GFP-mESCs), red fluorescent protein (RFP-mESCs), and cyan fluorescent protein (CFP-mESCs) into wild-type blastocysts (Ueno and Weissman, 2006). Within tetrachimera kidneys, mature nephrons were polyclonal, revealing mixed contributions of clones to individual tubule segments (Fig- ures "
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    • "In mice, the primitive phase of hematopoiesis occurs in the yolk sac at embryonic (E) day 7.5 [2] [3] [4] [5]. It results in the production of primitive hematopoietic cells including mainly large, nucleated erythroblasts, some megakaryocytes, and primitive macrophages needed for embryonic growth [2] [3] [4] [5]. The definitive phase of hematopoiesis in mice begins in the extraembryonic yolk sac at E8.25 [2] [6]. "

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