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Cell cycle distribution of cord blood‐derived haematopoietic progenitor cells and their recruitment into the S‐phase of the cell cycle
Author(s) -
Claudia Lucotti,
Lucia Malabarba,
Vittorio Rosti,
Gaetano Bergamaschi,
Marco Danova,
Rosangela Invernizzi,
Alessandro Pecci,
Isabella Ramajoli,
Cesare Perotti,
Lorella Torretta,
Mara De Amici,
L. Salvaneschi,
Mario Cazzola
Publication year - 2000
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.2000.01915.x
Subject(s) - cell cycle , haematopoiesis , stem cell factor , progenitor cell , biology , microbiology and biotechnology , interleukin 3 , flow cytometry , cd34 , cord blood , stem cell , cell culture , immunology , cell , t cell , biochemistry , antigen presenting cell , genetics , immune system
The objective of this study was to evaluate the cycling status of cord blood (CB)‐derived colony‐forming cells (CFC) and long‐term culture‐initiating cells (LTC‐IC), and their recruitment into the S‐phase of the cell cycle. By using the cytosine arabinoside (Ara‐C) suicide approach, we found that only small proportions of both CFC and LTC‐IC were in the S‐phase of the cell cycle. These estimates were confirmed by flow cytometric DNA analysis, which showed that 96 ± 2% of CB‐derived CD34+ cells were in G 0 /G 1 and only 1.6 ± 0.4% in the S‐phase. Staining of CD34+ cells with an antistatin monoclonal antibody, a marker of the G 0 phase, indicated that among CD34+ cells with a flow cytometric DNA content typical of the G 0 /G 1 phase 68 ± 7% of cells were in the G 0 phase of the cell cycle. Incubation (24 h) with interleukin 3 (IL‐3), recombinant human stem cell factor (SCF) and granulocyte colony‐stimulating factor (G‐CSF) significantly increased the proportion of cells in the S‐phase for both CFC and LTC‐IC without inducing any loss in numbers. Flow cytometric DNA analysis also showed an increase in CD34+ cells in the S‐phase upon continuous exposure to these cytokines. Our findings indicate that: (i) very few CB‐derived CFC or LTC‐IC were in the S‐phase of the cell cycle; (ii) a substantial amount of CD34+ cells with a flow cytometric DNA content typical of the G 0 /G 1 fraction was cycling, as found in the G 1 phase of the cell cycle; and (iii) 24‐h incubation with IL‐3, SCF and G‐CSF could drive a proportion of progenitor cells into the S‐phase without reducing their number. These data might be useful for gene transfer protocols and the ex vivo expansion of CB‐derived progenitor cells.