Hematopoietic stem cells in co-culture with mesenchymal stromal cells -modeling the niche compartments in vitro

Duohui Jing, Ana Violeta Fonseca, Nael Alakel, Fernando A Fierro, Katrin Muller, Martin Bornhauser, Gerhard Ehninger, Denis Corbeil, Rainer Ordemann

Research output: Contribution to journalArticle

141 Scopus citations

Abstract

Background: Hematopoietic stem cells located in the bone marrow interact with a specific microenvironment referred to as the stem cell niche. Data derived from ex vivo co-culture systems using mes-enchymal stromal cells as a feeder cell layer suggest that cell-to-cell contact has a significant impact on the expansion, migratory potential and 'stemness' of hematopoietic stem cells. Here we investigated in detail the spatial relationship between hematopoietic stem cells and mes-enchymal stromal cells during ex vivo expansion. Design and Methods: In the co-culture system, we defined three distinct localizations of hematopoietic stem cells relative to the mesenchymal stromal cell layer: (i) those in supernatant (non-adherent cells); (ii) those adhering to the surface of mesenchymal stromal cells (phase-bright cells) and (iii) those beneath the mesenchymal stromal cells (phase-dim cells). Cell cycle, proliferation, cell division and immunophenotype of these three cell fractions were evaluated from day 1 to 7. Results: Phase-bright cells contained the highest proportion of cycling progenitors during co-culture. In contrast, phase-dim cells divided much more slowly and retained a more immature phenotype compared to the other cell fractions. The phase-dim compartment was soon enriched for CD34+/CD38-cells. Migration beneath the mesenchymal stromal cell layer could be hampered by inhibiting integrin ß1 or CXCR4. Conclusions: Our data suggest that the mesenchymal stromal cell surface is the predominant site of prolifer-ation of hematopoietic stem cells, whereas the compartment beneath the mesenchymal stro-mal cell layer seems to mimic the stem cell niche for more immature cells. The SDF-1/CXCR4 interaction and integrin-mediated cell adhesion play important roles in the distribution of hematopoietic stem cells in the co-culture system.

Original languageEnglish (US)
Pages (from-to)542-550
Number of pages9
JournalHaematologica
Volume95
Issue number4
DOIs
StatePublished - Apr 2010
Externally publishedYes

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Keywords

  • CXCR4
  • Hematopoietic stem cells
  • Integrin β1

ASJC Scopus subject areas

  • Hematology

Cite this

Jing, D., Fonseca, A. V., Alakel, N., Fierro, F. A., Muller, K., Bornhauser, M., Ehninger, G., Corbeil, D., & Ordemann, R. (2010). Hematopoietic stem cells in co-culture with mesenchymal stromal cells -modeling the niche compartments in vitro. Haematologica, 95(4), 542-550. https://doi.org/10.3324/haematol.2009.010736