Erythropoietic differentiation of a human embryonic stem cell line harbouring the sickle cell anaemia mutation.
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.
Pryzhkova
Marina V
MV
Institute for Cell Engineering, Stem Cell Program, Johns Hopkins University School of Medicine, 733 N Broadway, BRB 755 Baltimore, MD 21205, USA.
Peters
Ann
A
Zambidis
Elias T
ET
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
D016895:Culture Media, Serum-Free; D017931:DNA Primers