Ex vivo expansion of mature human neutrophils with normal functions from purified peripheral blood CD34+ haematopoietic progenitor cells.
Purified CD34+ haematopoietic progenitor cells were cultivated with stem cell factor, interleukin 3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte CSF (G-CSF) for 7 d, and thereafter non-adherent cells were divided into two groups. Cells in one group (group A) were further cultivated for 7 d with four cytokines, and cells in the other group (group B) were further cultivated for 7 d with G-CSF alone. On day 14, 220-fold and 130-fold increases in the numbers of non-adherent cells were achieved for groups A and B respectively. These cell preparations contained 65% granulocytes for group A and 95% granulocytes for group B. These cells gained the ability to respond effectively with chemotaxis, phagocytosis and superoxide (O2-) release. Cells in group B were appropriately primed by G-CSF, GM-CSF, tumour necrosis factor alpha and IL-1beta for enhanced release of O2 -. The responsiveness of these cells was identical to that of peripheral blood neutrophils, indicating that cells in group B may be in the resting state. In contrast, cells in group A were not primed by these cytokines for enhanced release of O2- and released a large amount of O2- spontaneously, indicating that cells in group A may be in the activated state. These findings indicate that mature neutrophils with normal functions were expanded ex vivo in group B and suggest that these cells could be used for possible autologous neutrophil transfusion to prevent bacterial infections during severe neutropenia after cytotoxic chemotherapy.
Hino
M
M
Departments of Clinical Haematology and Physiology, Osaka City University Medical School, 1-5-7, Asahi-machi, Abeno-ku, Osaka 545-8586, Japan. hinom@med.osaka-cu.ac.jp
Suzuki
K
K
Yamane
T
T
Sakai
N
N
Kubota
H
H
Koh
K R
KR
Ohta
K
K
Hato
F
F
Kitagawa
S
S
Tatsumi
N
N
D018952:Antigens, CD34 / Q000276:immunology*; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D009504:Neutrophils / Q000166:cytology* / Q000637:transplantation; D011994:Recombinant Proteins / Q000494:pharmacology; D019089:Stem Cell Factor / Q000494:pharmacology
D018952:Antigens, CD34; D006133:Growth Substances; D007377:Interleukin-3; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor