Characterisation of Wnt gene expression during the differentiation of murine embryonic stem cells in vitro: role of Wnt3 in enhancing haematopoietic differentiation. Mech Dev 2001 May;103(1-2):49-59
Date
05/04/2001Pubmed ID
11335111DOI
10.1016/s0925-4773(01)00331-8Scopus ID
2-s2.0-0035018077 (requires institutional sign-in at Scopus site) 68 CitationsAbstract
The first haematopoietic stem cells in mammalian and non-mammalian vertebrates are derived from mesoderm, therefore genes that are important in mesoderm patterning and formation might also play an essential role in haematopoietic stem cell commitment and differentiation. Several members of the Wnt gene family are expressed in very specific patterns in embryonic mesoderm and have previously been shown to act as haematopoietic growth factors. In order to investigate in detail the role that such secreted proteins play in the biology of early haematopoietic commitment we have used in vitro differentiation of murine embryonal stem (ES) as a model system. Using reverse-transcriptase polymerase chain reaction analysis we identified several candidate Wnt genes whose expression pattern was consistent with a role in generation, maintenance and/or differentiation of early haematopoietic progenitor cells including three genes previously shown to have a role in haematopoiesis (Wnt5a, Wnt2b and Wnt10b). The most interesting candidate was Wnt3, because of its strong and regulated expression during in vitro differentiation of murine ES cells as well as its early embryonic expression in mesoderm. Overexpression of Wnt3 was sufficient to cause a consistent increase in the number of embryoid bodies committing to haematopoiesis further strengthening the evidence that this protein can enhance haematopoietic commitment during in vitro differentiation of ES cells. In addition, overexpression of Wnt3 caused a marked upregulation of Brachyury expression, thus providing some evidence that Brachyury may be one of the target genes for the Wnt3 signalling pathway.
Author List
Lako M, Lindsay S, Lincoln J, Cairns PM, Armstrong L, Hole NAuthor
Joy Lincoln PhD Professor in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsCOS Cells
Cell Differentiation
Cell Line
Cells, Cultured
Colony-Forming Units Assay
Dimethyl Sulfoxide
Embryo, Mammalian
Fetal Proteins
Green Fluorescent Proteins
Hematopoietic Stem Cells
Luminescent Proteins
Mesoderm
Mice
Microscopy, Fluorescence
Proto-Oncogene Proteins
Reverse Transcriptase Polymerase Chain Reaction
Signal Transduction
T-Box Domain Proteins
Time Factors
Transfection
Transformation, Genetic
Tretinoin
Up-Regulation
Wnt Proteins
Zebrafish Proteins