Functional and morphological characterization of immunomagnetically selected CD34+ hematopoietic progenitor cells

Federica Servida, Davide Soligo, Lorenza Caneva, Francesco Bertolini, Etienne De Harven, Simona Campiglio, Chiara Corsini, Giorgio Lambertenghi Deliliers

Research output: Contribution to journalArticlepeer-review

Abstract

We evaluated the potential of immunomagnetically selected (miniMACS) progenitor cells to give rise to colony-forming cells and their precursors, detected as long-term culture-initiating cells (LTC-IC), as well as their capacity to expand in liquid cultures. A 90% mean purity, a 43.2% yield and a 55.8-fold enrichment were achieved from normal bone marrow. When corrected for enrichment, the mean number of committed progenitor cells and the frequency of LTC-IC (evaluated by means of limiting dilution assay [LDA]) were not statistically different in low density mononuclear cells or in the CD34-enriched fractions. In five cases CD34+ selected cells grown in a stroma-free long-term bone marrow culture system with the addition of stem cell factor, interleukin 3, interleukin 6 and GM-CSF every 48 h, showed a 15 (±15) and 31 (±21) mean colony forming unit-granulocyte/macrophage fold increase on cultures at days 7 and 14. However, when corrected for enrichment, the expansion capability of these cells was significantly lower than that of the unseparated fraction, particularly after the first week. Immediately after separation, electron microscopy revealed that the CD34+ selected fraction contained more than 45% of well-differentiated myeloid cells (MPO+), with iron heads preferentially clustered at one pole of the cell surface and sometimes already endocytosed in pinocytic vesicles. After 24 h and 48 h incubation at 37°C, the majority of the cells showed no iron particles, but about 30% of the cells were iron-labeled phagocytic cells. The percentage of apoptotic cells with internalized iron was negligible. These data show that immunomagnetically separated CD34+ cells may have a slightly impaired short-term expansion capability, but give rise to cells. During the separation, the iron beads are internalized, rapidly processed in the cytoplasm and do not seem to interfere with in vitro growth.

Original languageEnglish
Pages (from-to)430-438
Number of pages9
JournalStem Cells
Volume14
Issue number4
Publication statusPublished - Jul 1996

Keywords

  • CD34 antigen
  • Cell separation
  • Ex vivo expansion
  • Growth factors
  • Hematopoietic stem cells
  • Long-term culture-initiating cells
  • Ultrastructure

ASJC Scopus subject areas

  • Cell Biology

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