title: Induction of hematopoietic differentiation of mouse embryonic stem cells by an AGM-derived stromal cell line is not further enhanced by overexpression of HOXB4.
abstract: Hematopoietic differentiation of embryonic stem (ES) cells can be enhanced by co-culture with stromal cells derived from hematopoietic tissues and by overexpression of the transcription factor HOXB4. In this study, we compare the hematopoietic inductive effects of stromal cell lines derived from different subregions of the embryonic aorta-gonad-mesonephros tissue with the commonly used OP9 stromal cell line and with HOXB4 activation. We show that stromal cell lines derived from the aorta and surrounding mesenchyme (AM) act at an earlier stage of the differentiation process compared with the commonly used OP9 stromal cells. AM stromal cells were able to promote the further differentiation of isolated brachyury-GFP(+) mesodermal cells into hematopoietic progenitors, whereas the OP9 stromal cells could not support the differentiation of these cells. Co-culture and analyses of individual embryoid bodies support the hypothesis that the AM stromal cell lines could enhance the de novo production of hematopoietic progenitors, lending support to the idea that AM stromal cells might act on prehematopoietic mesoderm. The induction level observed for AM stromal cells was comparable to HOXB4 activation, but no additive effect was observed when these 2 inductive strategies were combined. Addition of a γ-secretase inhibitor reduced the inductive effects of both the stromal cell line and HOXB4, providing clues to possible shared molecular mechanisms.
authors.affiliation: MRC Centre for Regenerative Medicine, John Hughes Bennett Laboratory, Queens Medical Research Institute, Edinburgh, United Kingdom.
authors.forename: Sabrina A M
authors.initials: SA
authors.lastname: Gordon-Keylock
authors.forename: Melany
authors.initials: M
authors.lastname: Jackson
authors.forename: Caoxin
authors.initials: C
authors.lastname: Huang
authors.forename: Kay
authors.initials: K
authors.lastname: Samuel
authors.forename: Richard A
authors.initials: RA
authors.lastname: Axton
authors.forename: Robert A J
authors.initials: RA
authors.lastname: Oostendorp
authors.forename: Helen
authors.initials: H
authors.lastname: Taylor
authors.forename: Julie
authors.initials: J
authors.lastname: Wilson
authors.forename: Lesley M
authors.initials: LM
authors.lastname: Forrester
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors; D000818:Animals; D001011:Aorta / Q000166:cytology* / Q000196:embryology; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D018920:Coculture Techniques; D004622:Embryo, Mammalian / Q000166:cytology; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D005326:Fetal Proteins / Q000235:genetics / Q000378:metabolism; D006410:Hematopoiesis / Q000502:physiology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008648:Mesoderm / Q000166:cytology* / Q000196:embryology; D008650:Mesonephros / Q000166:cytology / Q000196:embryology; D051379:Mice; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D000098908:Brachyury Protein
chemical_list: D053829:Amyloid Precursor Protein Secretases; D015415:Biomarkers; D005326:Fetal Proteins; D018398:Homeodomain Proteins; D020825:T-Box Domain Proteins; D014157:Transcription Factors; D000098908:Brachyury Protein; C513262:Hoxb4 protein, mouse