Angiopoietin-like 5 and IGFBP2 stimulate ex vivo expansion of human cord blood hematopoietic stem cells as assayed by NOD/SCID transplantation.
Hematopoietic stem cells (HSCs) are the basis of bone marrow transplantation and are attractive target cells for hematopoietic gene therapy, but these important clinical applications have been severely hampered by difficulties in ex vivo expansion of HSCs. In particular, the use of cord blood for adult transplantation is greatly limited by the number of HSCs. Previously we identified angiopoietin-like proteins and IGF-binding protein 2 (IGFBP2) as new hormones that, together with other factors, can expand mouse bone marrow HSCs in culture. Here, we measure the activity of multipotent human severe combined immunodeficient (SCID)-repopulating cells (SRCs) by transplantation into the nonobese diabetic SCID (NOD/SCID) mice; secondary transplantation was performed to evaluate the self-renewal potential of SRCs. A serum-free medium containing SCF, TPO, and FGF-1 or Flt3-L cannot significantly support expansion of the SRCs present in human cord blood CD133+ cells. Addition of either angiopoietin-like 5 or IGF-binding protein 2 to the cultures led to a sizable expansion of HSC numbers, as assayed by NOD/SCID transplantation. A serum-free culture containing SCF, TPO, FGF-1, angiopoietin-like 5, and IGFBP2 supports an approximately 20-fold net expansion of repopulating human cord blood HSCs, a number potentially applicable to several clinical processes including HSC transplantation.
Zhang
Cheng Cheng
CC
Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA. alec.zhang@utsouthwestern.edu
Kaba
Megan
M
Iizuka
Satoru
S
Huynh
HoangDinh
H
Lodish
Harvey F
HF
D000071916:AC133 Antigen; D000075802:Angiopoietin-like Proteins; D042682:Angiopoietins; D000818:Animals; D015703:Antigens, CD; D001324:Autoantigens / Q000494:pharmacology; D018929:Cell Culture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology; D015316:Genetic Therapy; D006023:Glycoproteins; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D018971:Insulin-Like Growth Factor Binding Protein 2 / Q000494:pharmacology*; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D007453:Iodide Peroxidase / Q000494:pharmacology; D033862:Iron-Binding Proteins / Q000494:pharmacology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D010455:Peptides; D014183:Transplantation, Heterologous
D000071916:AC133 Antigen; C473567:ANGPTL5 protein, human; D000075802:Angiopoietin-like Proteins; D042682:Angiopoietins; D015703:Antigens, CD; D001324:Autoantigens; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D006023:Glycoproteins; D018971:Insulin-Like Growth Factor Binding Protein 2; D036341:Intercellular Signaling Peptides and Proteins; D033862:Iron-Binding Proteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D016220:Fibroblast Growth Factor 1; C056658:TPO protein, human; D007453:Iodide Peroxidase