Development of a vascular niche platform for expansion of repopulating human cord blood stem and progenitor cells.
Transplantation of ex vivo expanded human umbilical cord blood cells (hCB) only partially enhances the hematopoietic recovery after myelosuppressive therapy. Incubation of hCB with optimal combinations of cytokines and niche cells, such as endothelial cells (ECs), could augment the efficiency of hCB expansion. We have devised an approach to cultivate primary human ECs (hECs) in serum-free culture conditions. We demonstrate that coculture of CD34(+) hCB in direct cellular contact with hECs and minimal concentrations of thrombopoietin/Kit-ligand/Flt3-ligand resulted in a 400-fold expansion of total hematopoietic cells, 150-fold expansion of CD45(+)CD34(+) progenitor cells, and 23-fold expansion of CD45(+) Lin(-)CD34(hi+)CD45RA(-)CD49f(+) stem and progenitor cells over a 12-day period. Compared with cytokines alone, coculture of hCB with hECs permitted greater expansion of cells capable of multilineage engraftment and serial transplantation, hallmarks of long-term repopulating hematopoietic stem cells. Therefore, hECs establish a cellular platform for expansion of hematopoietic stem and progenitor cells and treatment of hematologic disorders.
Department of Genetic Medicine, Weill Cornell Medical College, New York, NY 10065, USA. jmb2009@med.cornell.edu
Jason M
JM
Butler
Eric J
EJ
Gars
Daylon J
DJ
James
Daniel J
DJ
Nolan
Joseph M
JM
Scandura
Shahin
S
Rafii
D000818:Animals; D001808:Blood Vessels / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D002478:Cells, Cultured; D036101:Cord Blood Stem Cell Transplantation / Q000379:methods; D005312:Fetal Blood / Q000166:cytology* / Q000502:physiology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D008822:Mice, Transgenic; D055153:Stem Cell Niche / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds