title: Erythropoietin-stimulated endothelial cells support erythroid cell differentiation of CD34(+) haematopoietic progenitors.
abstract: OBJECTIVE
  Endothelial cells provide a unique medium for the proliferation and white lineage differentiation of haematopoietic progenitor cells (HPC). Whether this quality can be exploited to facilitate the differentiation of erythroid precursors is not yet known.
  
  
  METHODS
  Haematopoietic progenitor cells derived from cord blood were cultured for 3 weeks in erythropoietin-stimulated supernatants with (n = 6) or without cyclosporine A (CSA, n = 6). Cell count, phenotype and morphology were assessed on a weekly basis, and the haemoglobin content was analysed. These cultures were compared with erythroid differentiation induced by cytokines only (n = 6).
  
  
  RESULTS
  Endothelial supernatants combined with CSA led to equivalent numbers of CD71(+) erythroblasts after 1 week as cytokines only. The purity of glycophorin-positive, CD45-negative cells was higher in cells generated in endothelial supernatants than in cytokine-based media. Additional prostaglandin E2 induced a change from fetal to adult haemoglobin.
  
  
  CONCLUSIONS
  For the generation of erythroblasts from HPC, endothelial supernatants are a simple and cost-effective alternative to culture conditions based on cytokines.
authors.affiliation: Institute for Transfusion Medicine, Charité - Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Zaid
authors.forename: C
authors.initials: C
authors.lastname: Frömmel
authors.forename: A
authors.initials: A
authors.lastname: Lun
authors.forename: A
authors.initials: A
authors.lastname: Moldenhauer
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D016572:Cyclosporine / Q000494:pharmacology; D016207:Cytokines / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D004900:Erythroblasts / Q000166:cytology*; D041905:Erythroid Cells / Q000166:cytology*; D004920:Erythropoiesis; D004921:Erythropoietin / Q000494:pharmacology*; D005312:Fetal Blood / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D019089:Stem Cell Factor / Q000494:pharmacology
chemical_list: D018952:Antigens, CD34; D016207:Cytokines; D019089:Stem Cell Factor; D004921:Erythropoietin; D016572:Cyclosporine
keywords: blood components; cell culture; differentiation; hematopoietic stem cell; transfusion therapy