A human stromal-based serum-free culture system supports the ex vivo expansion/maintenance of bone marrow and cord blood hematopoietic stem/progenitor cells.
OBJECTIVE
We investigated the role of human stromal layers (hu-ST) on the ex vivo expansion/maintenance of human hematopoietic stem/progenitor cells (HSC) from adult bone marrow (BM) and umbilical cord blood (CB).


METHODS
BM and CB CD34(+)-enriched cells were cultured in serum-free medium supplemented with SCF, bFGF, LIF, and Flt-3, in the presence or absence of stroma, and analyzed for proliferation, phenotype, and clonogenic potential.


RESULTS
Significant expansion of BM and CB CD34(+) and CD34(+)CD38(-) cells were achieved in the presence of hu-ST. The differentiative potential of both BM and CB CD34(+)-enriched cells cocultured with hu-ST was primarily shifted toward the myeloid lineage, while maintaining/expanding a CD7(+) population. Clonogenic analysis of the expanded cells showed increases in progenitors of the myeloid lineage, including colony-forming unit-granulocyte, macrophage (CFU-GM) and colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte (CFU-Mix) for both BM (stroma and stroma-free conditions) and CB cells in the presence of stroma.


CONCLUSIONS
These results indicate that adult hu-ST in the presence of appropriate cytokines can be used to efficiently expand/maintain myeloid and lymphoid cell populations from human BM and CB HSC.
da Silva
Cláudia Lobato
CL
Department of Animal Biotechnology, University of Nevada, Reno, 89557, USA.
Gonçalves
Raquel
R
Crapnell
Kirsten B
KB
Cabral
Joaquim M S
JM
Zanjani
Esmail D
ED
Almeida-Porada
Graça
G
D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000097:blood; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000502:physiology*; D016895:Culture Media, Serum-Free; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007231:Infant, Newborn; D007700:Kinetics; D017154:Stromal Cells / Q000166:cytology* / Q000502:physiology; D013997:Time Factors
D015703:Antigens, CD; D018952:Antigens, CD34; D016895:Culture Media, Serum-Free