title: Erythropoietic differentiation of a human embryonic stem cell line harbouring the sickle cell anaemia mutation.
abstract: Herein is reported efficient erythropoietic differentiation of a human embryonic stem cell (ESC) line derived from a preimplantation genetic diagnosis (PGD)-screened embryo that harbours the homozygous sickle cell disease (SCD) haemoglobinopathy mutation. This human ESC line possesses typical pluripotency characteristics and forms multilineage teratomas in vivo. SCD-human ESC efficiently differentiated to the haematopoietic lineage under serum-free and stromal co-culture conditions and gave rise to robust primitive and definitive erythrocytes. Expression of embryonic, fetal and adult sickle globin genes in SCD PGD-derived human ESC-derived erythrocytes was confirmed by quantitative real-time PCR, intracytoplasmic fluorescence-activated cell sorting and in-situ immunostaining of PGD-derived human ESC teratoma sections. These data introduce important methodologies and paradigms for using patient-specific human ESC to generate normal and haemoglobinopathic erythroid progenitors for biomedical research.
authors.affiliation: Institute for Cell Engineering, Stem Cell Program, Johns Hopkins University School of Medicine, 733 N Broadway, BRB 755 Baltimore, MD 21205, USA.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Pryzhkova
authors.forename: Ann
authors.initials: A
authors.lastname: Peters
authors.forename: Elias T
authors.initials: ET
authors.lastname: Zambidis
mesh_terms: D000755:Anemia, Sickle Cell / Q000235:genetics*; D001483:Base Sequence; D018920:Coculture Techniques; D016895:Culture Media, Serum-Free; D017931:DNA Primers; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D007621:Karyotyping; D009154:Mutation; D016133:Polymerase Chain Reaction
chemical_list: D016895:Culture Media, Serum-Free; D017931:DNA Primers