Expansion of mobilized peripheral blood progenitor cells under defined culture conditions rsing CD34+CD71-CD45- cells as a starting population.
A major goal of experimental and clinical hematology is the identification of mechanisms and conditions supporting the expansion of transplantable hematopoietic stem cells. We assessed the expansion potential of CD34+CD71-CD45- cells derived from granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood under recently defined serum-free culture conditions. The CD34+CD71-CD45- cells in mobilized peripheral blood were found to contain the majority (92%+/-5.6) of primitive long-term culture initiating cells (LTCIC) and 53.5%+/-16.7 of the more committed colony-forming cells (CFC). Furthermore, this population represents 23.3%+/-4.1 of the total CD34+ cells and allows reduction of the cell density important for maintenance/expansion of primitive progenitor cells. CD34+ CD71- CD45- cells were cultured in defined serum-free media supplemented with 300 ng each of Flt-3 ligand and stem cell factor (SCF), 60 ng of interleukin (IL)-3, and 20 ng each of IL-6 and G-CSF. Mononuclear cells (MNC) and CFC were expanded 50-fold and 200-fold, respectively; primitive progenitor cells (LTC-IC) were maintained at input values after a total of 10 days of expansion. The addition of IL-15 to our cytokine cocktail expanded LTC-IC 2- to 3-fold and CFC to >500-fold. The data presented should allow clinical manipulation (purging) and expansion procedures with mobilized PBPC harvests without the loss of primitive progenitor cells and could be made applicable for large-scale clinical expansion.
Zech
Nicolas H
NH
Stem Cell Laboratory, Division of Hematology and Oncology, University Hospital Innsbruck, Innsbruck, Austria.
Gunsilius
Eberhard
E
Clausen
Johannes
J
Hoflehner
Elisabeth
E
Hoelzler
Karoline
K
Gastl
Günther
G
Petzer
Andreas L
AL
D036541:ADP-ribosyl Cyclase / Q000096:biosynthesis; D051997:ADP-ribosyl Cyclase 1; D015703:Antigens, CD / Q000096:biosynthesis*; D018952:Antigens, CD34 / Q000096:biosynthesis*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000096:biosynthesis*; D001943:Breast Neoplasms / Q000097:blood; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004921:Erythropoietin / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D019409:Interleukin-15 / Q000378:metabolism; D007377:Interleukin-3 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000096:biosynthesis*; D008228:Lymphoma, Non-Hodgkin / Q000097:blood; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000378:metabolism; D008747:Methylcellulose / Q000737:chemistry; D011990:Receptors, Transferrin; D013234:Stem Cells / Q000378:metabolism*
D015703:Antigens, CD; D018952:Antigens, CD34; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D016895:Culture Media, Serum-Free; D019409:Interleukin-15; D007377:Interleukin-3; D015850:Interleukin-6; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011990:Receptors, Transferrin; C084881:flt3 ligand protein; D004921:Erythropoietin; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D008747:Methylcellulose; D017493:Leukocyte Common Antigens; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1