Marrow cells surviving one week in vitro incubation with 5-fluorouracil (S-FU) in the presence of kit ligand (KL) and interleuldn-3 (TL-3) are candidate eady progenitor cells (Berardi et ai Science 267,104,1995). Aim of the study: to determine the phenotype and the functional properties of cord blood surviving 5-FU treatment. Methods: monnuclear cells from 17 cord blood obtained after ficoll separation were incubated with KL(50 ng/ml) and IL-3 (100 ng/ml) in the presence of 5-FU (600 ng/ml). After one week the phenotype and the cell cycle status (BrdU posinvity) of the remaining cells were determined as well as their capacity to give rise to CFU-GM either directly either after 5 week stromal cell culture (LTC-IC). Results: before incubation with 5-FU: mean cell number=96×106, CD34+=24×104, CD34+/38-=2.5×104, BrdU=15.3×106, CFU-GM=6.9×104 after incubation with 5-FU: mean cell number=15.6×104, CD34+=6.8×104, CD34+/38-=0.35×104, BrdU=0.15x10, CFU-OM=0 but 12.6 five weeks CFU-GM output. KL plays a crucial role in the maintenance of early progenitors as suggested by: a) its role in the maintenance viable cells, the high percentage of cells (35.S) and CD34+ cells (75.5) expressing c-kit after 5-FU treatment; b) the fact that it increase the CFU-GM production (5.3 fold) when added to stromal cell cultures, c) the abrogation of LTC-IC growth by the addition of Ab antic-kit. Conclusion: our study demonstrates that cord blood is rich in early progenitor cells, indicates the crucial role of KL in maintaining early progenitors and suggests the capacity of these progenitors to sustain the hcmopoiesis for a long period (i.e. several years after transplantation).
Characterization of hematopoietic stem cells from human cord blood
Fagioli F.;Berger M.;Basso G.;Sciascia C.;Sanavio F.
1997-01-01
Abstract
Marrow cells surviving one week in vitro incubation with 5-fluorouracil (S-FU) in the presence of kit ligand (KL) and interleuldn-3 (TL-3) are candidate eady progenitor cells (Berardi et ai Science 267,104,1995). Aim of the study: to determine the phenotype and the functional properties of cord blood surviving 5-FU treatment. Methods: monnuclear cells from 17 cord blood obtained after ficoll separation were incubated with KL(50 ng/ml) and IL-3 (100 ng/ml) in the presence of 5-FU (600 ng/ml). After one week the phenotype and the cell cycle status (BrdU posinvity) of the remaining cells were determined as well as their capacity to give rise to CFU-GM either directly either after 5 week stromal cell culture (LTC-IC). Results: before incubation with 5-FU: mean cell number=96×106, CD34+=24×104, CD34+/38-=2.5×104, BrdU=15.3×106, CFU-GM=6.9×104 after incubation with 5-FU: mean cell number=15.6×104, CD34+=6.8×104, CD34+/38-=0.35×104, BrdU=0.15x10, CFU-OM=0 but 12.6 five weeks CFU-GM output. KL plays a crucial role in the maintenance of early progenitors as suggested by: a) its role in the maintenance viable cells, the high percentage of cells (35.S) and CD34+ cells (75.5) expressing c-kit after 5-FU treatment; b) the fact that it increase the CFU-GM production (5.3 fold) when added to stromal cell cultures, c) the abrogation of LTC-IC growth by the addition of Ab antic-kit. Conclusion: our study demonstrates that cord blood is rich in early progenitor cells, indicates the crucial role of KL in maintaining early progenitors and suggests the capacity of these progenitors to sustain the hcmopoiesis for a long period (i.e. several years after transplantation).
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