Extensive amplification and self-renewal of human primitive hematopoietic stem cells from cord blood.
The use of umbilical cord blood as a source of marrow repopulating cells for the treatment of pediatric malignancies has been established. Given the general availability, the ease of procurement, and progenitor content, cord blood is an attractive alternative to bone marrow or growth factor mobilized peripheral blood cells as a source of transplantable hematopoietic tissue. However, there is a major potential limitation to the widespread use of cord blood as a source of hematopoietic stem cells for marrow replacement and gene therapy. There may be enough hematopoietic stem cells to reconstitute children, but the ability to engraft an adult might require ex vivo manipulations. We describe an in vitro system in which the growth of cord blood CD34+ cells is sustained and greatly expanded for more than 6 months by the simple combination of two hematopoietic growth factors. Progenitors and cells belonging to all hematopoietic lineages are continuously and increasingly generated (the number of colony-forming unit-granulocyte-macrophage [CFU-GM] present at the end of 6 months of culture are well over 2,000,000-fold the CFU-GM present at the beginning of the culture). Very primitive hematopoietic progenitors, including long-term culture-initiating cells (LTC-ICs) and blast cell colony-forming units, are also greatly expanded (after 20 weeks of liquid culture, LTC-IC number is over 200,000-fold the initial number). The extremely prolonged maintenance and the massive expansion of these progenitors, which share many similarities with murine long-term repopulating cells, suggest that extensive renewal and little differentiation take place. This system might prove useful in diverse clinical settings involving treatment of grown-up children and adults with transplantation of normal or genetically manipulated hematopoietic stem cells.
Piacibello
W
W
Department of Biomedical Sciences, Torino Medical School, University of Torino, Italy.
Sanavio
F
F
Garetto
L
L
Severino
A
A
Bergandi
D
D
Ferrario
J
J
Fagioli
F
F
Berger
M
M
Aglietta
M
M
D018952:Antigens, CD34 / Q000032:analysis; D002469:Cell Separation; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D005312:Fetal Blood / Q000166:cytology*; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D007231:Infant, Newborn; D008565:Membrane Proteins / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology; D013926:Thrombopoietin / Q000494:pharmacology*
D018952:Antigens, CD34; D016298:Hematopoietic Cell Growth Factors; D008565:Membrane Proteins; D011994:Recombinant Proteins; C084881:flt3 ligand protein; D013926:Thrombopoietin