title: In vitro expansion of human cord blood CD36+ erythroid progenitors: temporal changes in gene and protein expression.
abstract: OBJECTIVE
  Erythropoiesis involves proliferation and differentiation of committed erythroid progenitors to mature red blood cells. The objective of this study was to characterize growth characteristics of human CD36+ erythroid progenitors and to profile temporal expression of lineage-specific transcription factors, structural proteins, and growth factor receptors involved in erythropoiesis.
  
  
  METHODS
  Erythropoietin-induced differentiation of human cord blood CD36+ erythroid progenitors was profiled for GATA-1, GATA-2, NFE2, EKLF, SCL, PU.1, Id1, Evi-1, c-myb, Hox2.2, c-kit, EpoR, glycophorin A (GPA), CD71, beta- and gamma-globin, and protein 4.2 gene and/or protein expression and DNA content analysis on days 4, 7, and 15 of culture.
  
  
  RESULTS
  Real-time RT-PCR analysis revealed upregulation of GATA-1, Id1, glycophorin A, and protein 4.2 mRNA expression on day 7 when compared to day 4 and decreased expression on day 15. EKLF, GATA-2, Hox2.2, c-myb, Evi-1, c-kit, and PU.1 mRNA expression decreased on days 7 and 15. NFE2, CD71, SCL, and EPO-R mRNA expression remained similar on days 4 and 7 but decreased on day 15. Expression of globin genes beta- and gamma-globin increased on both day 7 and day 15 compared to day 4. Values from flow cytometric quantitation of glycophorin A, transferrin receptor (CD71), and hemoglobin A proteins correlated with gene expression results. DNA analysis demonstrated that most cells lacked DNA content by day 15, a finding consistent with enucleation and terminal erythroid differentiation.
  
  
  CONCLUSIONS
  These data indicate that in vitro liquid cultures of committed CD36+ erythroid progenitor cells retain, in part, many features of erythropoiesis at the cellular and molecular level and may provide a useful model for assessment of disease-related or drug-induced erythropoietic abnormalities.
authors.affiliation: GlaxoSmithKline, Safety Assessment Department, King of Prussia, Pa. 19406, USA. Marshall.2.Scicchitano@gsk.com
authors.forename: Marshall S
authors.initials: MS
authors.lastname: Scicchitano
authors.forename: David C
authors.initials: DC
authors.lastname: McFarland
authors.forename: Lauren A
authors.initials: LA
authors.lastname: Tierney
authors.forename: Padma K
authors.initials: PK
authors.lastname: Narayanan
authors.forename: Lester W
authors.initials: LW
authors.lastname: Schwartz
mesh_terms: D018955:CD36 Antigens; D002454:Cell Differentiation; D002455:Cell Division; D019070:Cell Lineage / Q000235:genetics; D002478:Cells, Cultured; D015672:Erythroid Precursor Cells / Q000166:cytology* / Q000502:physiology*; D004920:Erythropoiesis / Q000235:genetics; D005434:Flow Cytometry; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics
chemical_list: D018955:CD36 Antigens; D011506:Proteins