Stepwise development of hematopoietic stem cells from embryonic stem cells

• Published in: PloS one Vol. 4; no. 3; p. e4820
• Main Authors: Matsumoto, Kenji, Isagawa, Takayuki, Nishimura, Toshinobu, Ogaeri, Takunori, Eto, Koji, Miyazaki, Satsuki, Miyazaki, Jun-ichi, Aburatani, Hiroyuki, Nakauchi, Hiromitsu, Ema, Hideo
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 16.03.2009; Public Library of Science (PLoS)
• Subjects: ABC transporters; Adenoviruses; Angiogenesis; Animals; c-Kit protein; CD45 antigen; Cell culture; Cell Differentiation; Cloning; Developmental Biology/Cell Differentiation; Developmental Biology/Embryology; Developmental Biology/Stem Cells; Developmental stages; Divergence; Embryo cells; Embryogenesis; Embryonic stem cells; Embryonic Stem Cells - cytology; Embryos; Fibroblasts; Flow cytometry; Gene Expression; Genomes; Hemangioblasts; Hematology/Hematopoiesis; Hematopoietic stem cells; Hematopoietic Stem Cells - cytology; HOXB4 protein; Medicine; Mice; Ontogeny; Population; Reverse Transcriptase Polymerase Chain Reaction; Rodents; Science; Stem cell transplantation; Stem cells; Stromal cells
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0004820

The cellular ontogeny of hematopoietic stem cells (HSCs) remains poorly understood because their isolation from and their identification in early developing small embryos are difficult. We attempted to dissect early developmental stages of HSCs using an in vitro mouse embryonic stem cell (ESC) differentiation system combined with inducible HOXB4 expression. Here we report the identification of pre-HSCs and an embryonic type of HSCs (embryonic HSCs) as intermediate cells between ESCs and HSCs. Both pre-HSCs and embryonic HSCs were isolated by their c-Kit(+)CD41(+)CD45(-) phenotype. Pre-HSCs did not engraft in irradiated adult mice. After co-culture with OP9 stromal cells and conditional expression of HOXB4, pre-HSCs gave rise to embryonic HSCs capable of engraftment and long-term reconstitution in irradiated adult mice. Blast colony assays revealed that most hemangioblast activity was detected apart from the pre-HSC population, implying the early divergence of pre-HSCs from hemangioblasts. Gene expression profiling suggests that a particular set of transcripts closely associated with adult HSCs is involved in the transition of pre-HSC to embryonic HSCs. We propose an HSC developmental model in which pre-HSCs and embryonic HSCs sequentially give rise to adult types of HSCs in a stepwise manner.