title: Effect of iron deficiency on c-kit⁺ cardiac stem cells in vitro.
abstract: OBJECTIVE
  Iron deficiency is a common comorbidity in chronic heart failure (CHF) which may exacerbate CHF. The c-kit⁺ cardiac stem cells (CSCs) play a vital role in cardiac function repair. However, much is unknown regarding the role of iron deficiency in regulating c-kit⁺ CSCs function. In this study, we investigated whether iron deficiency regulates c-kit⁺ CSCs proliferation, migration, apoptosis, and differentiation in vitro.
  
  
  METHODS
  All c-kit⁺ CSCs were isolated from adult C57BL/6 mice. The c-kit⁺ CSCs were cultured with deferoxamine (DFO, an iron chelator), mimosine (MIM, another iron chelator), or a complex of DFO and iron (Fe(III)), respectively. Cell migration was assayed using a 48-well chamber system. Proliferation, cell cycle, and apoptosis of c-kit⁺ CSCs were analyzed with BrdU labeling, population doubling time assay, CCK-8 assay, and flow cytometry. Caspase-3 protein level and activity were examined with Western blotting and spectrophotometric detection. The changes in the expression of cardiac-specific proteins (GATA-4,TNI, and β-MHC) and cell cycle-related proteins (cyclin D1, RB, and pRB) were detected with Western blotting.
  
  
  RESULTS
  DFO and MIM suppressed c-kit⁺ CSCs proliferation and differentiation. They also modulated cell cycle and cardiac-specific protein expression. Iron chelators down-regulated the expression and phosphorylation of cell cycle-related proteins. Iron reversed those suppressive effects of DFO. DFO and MIM didn't affect c-kit⁺ CSCs migration and apoptosis.
  
  
  CONCLUSIONS
  Iron deficiency suppressed proliferation and differentiation of c-kit⁺ CSCs. This may partly explain how iron deficiency affects CHF prognosis.
authors.affiliation: Department of Cardiology, Ninth People's Hospital, Shanghai Jiaotong University Medical School, Shanghai, PR China.
authors.forename: Dongqiang
authors.initials: D
authors.lastname: Song
authors.forename: Yuanmin
authors.initials: Y
authors.lastname: Li
authors.forename: Jiatian
authors.initials: J
authors.lastname: Cao
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Han
authors.forename: Lin
authors.initials: L
authors.lastname: Gao
authors.forename: Zuojun
authors.initials: Z
authors.lastname: Xu
authors.forename: Zhaofang
authors.initials: Z
authors.lastname: Yin
authors.forename: Guifang
authors.initials: G
authors.lastname: Wang
authors.forename: Yuqi
authors.initials: Y
authors.lastname: Fan
authors.forename: Changqian
authors.initials: C
authors.lastname: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003676:Deferoxamine / Q000494:pharmacology*; D005434:Flow Cytometry; D006321:Heart / Q000187:drug effects* / Q000502:physiology; D066298:In Vitro Techniques; D000090463:Iron Deficiencies; D051379:Mice; D008810:Mice, Inbred C57BL; D008898:Mimosine / Q000494:pharmacology*; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*; D017262:Siderophores / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
chemical_list: D017262:Siderophores; D008898:Mimosine; D019009:Proto-Oncogene Proteins c-kit; D053148:Caspase 3; D003676:Deferoxamine