title: Expansive effects of aorta-gonad-mesonephros-derived stromal cells on hematopoietic stem cells from embryonic stem cells.
abstract: BACKGROUND
  Hematopoietic stem cells (HSCs) give rise to all blood and immune cells and are used in clinical transplantation protocols to treat a wide variety of refractory diseases, but the amplification of HSCs has been difficult to achieve in vitro. In the present study, the expansive effects of aorta-gonad-mesonephros (AGM) region derived stromal cells on HSCs were explored, attempting to improve the efficiency of HSC transplantation in clinical practice.
  
  
  METHODS
  The murine stromal cells were isolated from the AGM region of 12 days postcoitum (dpc) murine embryos and bone marrow (BM) of 6 weeks old mice, respectively. After identification with flow cytometry and immunocytochemistry, the stromal cells were co-cultured with ESCs-derived, cytokines-induced HSCs. The maintenance and expansion of ESCs-derived HSCs were evaluated by detecting the population of CD34+ and CD34+Sca-1+ cells with flow cytometry and the blast colony-forming cells (BL-CFCs), high proliferative potential colony-forming cells (HPP-CFCs) by using semi-solid medium colonial culture. Finally, the homing and hematopoietic reconstruction abilities of HSCs were evaluated using a murine model of HSC transplantation in vivo.
  
  
  RESULTS
  AGM and BM-derived stromal cells were morphologically and phenotypically similar, and had the features of stromal cells. When co-cultured with AGM or BM stromal cells, more primitive progenitor cells (HPP-CFCs) could be detected in ESCs derived hematopoietic precursor cells, but BL-CFC's expansion could be detected only when co-cultured with AGM-derived stromal cells. The population of CD34+ hematopoietic stem/progenitor cells were expanded 3 times, but no significant expansion in the population of CD34+Sca-1+ cells was noted when co-cultured with BM stromal cells. While both CD34+ hematopoietic stem/progenitor cells and CD34+Sca-1+ cells were expanded 4 to 5 times respectively when co-cultured with AGM stromal cells. AGM region-derived stromal cells, like BM-derived stromal cells, could promote hematopoietic reconstruction and HSCs' homing to BM in vivo.
  
  
  CONCLUSIONS
  AGM-derived stromal cells in comparison with the BM-derived stromal cells could not only support the expansion of HSCs but also maintain the self-renewal and multi-lineage differentiation more effectively. They are promising in HSC transplantation.
authors.affiliation: Department of Hematology, Tongji Hospital, Tongji Medical School, Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Jin-rong
authors.initials: JR
authors.lastname: Fu
authors.forename: Wen-li
authors.initials: WL
authors.lastname: Liu
authors.forename: Yu-feng
authors.initials: YF
authors.lastname: Zhou
authors.forename: Jian-feng
authors.initials: JF
authors.lastname: Zhou
authors.forename: Han-ying
authors.initials: HY
authors.lastname: Sun
authors.forename: Li
authors.initials: L
authors.lastname: Luo
authors.forename: Heng
authors.initials: H
authors.lastname: Zhang
authors.forename: Hui-zhen
authors.initials: HZ
authors.lastname: Xu
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D001011:Aorta / Q000166:cytology*; D000067696:Ataxin-1; D000067528:Ataxins; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D004622:Embryo, Mammalian / Q000166:cytology*; D006066:Gonads / Q000166:cytology*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D008297:Male; D008650:Mesonephros / Q000166:cytology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009419:Nerve Tissue Proteins / Q000032:analysis; D009687:Nuclear Proteins / Q000032:analysis; D017154:Stromal Cells / Q000502:physiology*
chemical_list: D018952:Antigens, CD34; D000067696:Ataxin-1; D000067528:Ataxins; C000593750:Atxn1 protein, mouse; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins