Stable long-term blood formation by stem cells in murine steady-state hematopoiesis

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Oksana Zavidij - , German Cancer Research Center (DKFZ) (Author)
  • Claudia R. Ball - , National Center for Tumor Diseases (NCT) Heidelberg (Author)
  • Friederike Herbst - , German Cancer Research Center (DKFZ) (Author)
  • Felix Oppel - , German Cancer Research Center (DKFZ) (Author)
  • Sylvia Fessler - , German Cancer Research Center (DKFZ) (Author)
  • Manfred Schmidt - , German Cancer Research Center (DKFZ) (Author)
  • Christof Von Kalle - , German Cancer Research Center (DKFZ) (Author)
  • Hanno Glimm - , National Center for Tumor Diseases (NCT) Heidelberg (Author)

Abstract

Hematopoietic stem cells (HSCs) generate all mature blood cells during the whole lifespan of an individual. However, the clonal contribution of individual HSC and progenitor cells in steady-state hematopoiesis is poorly understood. To investigate the activity of HSCs under steady-state conditions, murine HSC and progenitor cells were genetically marked in vivo by integrating lentiviral vectors (LVs) encoding green fluorescent protein (GFP). Hematopoietic contribution of individual marked clones was monitored by determination of lentiviral integration sites using highly sensitive linear amplification- mediated-polymerase chain reaction. A remarkably stable small proportion of hematopoietic cells expressed GFP in LV-injected animals for up to 24 months, indicating stable marking of murine steady-state hematopoiesis. Analysis of the lentiviral integration sites revealed that multiple hematopoietic clones with both myeloid and lymphoid differentiation potential contributed to long-term hematopoiesis. In contrast to intrafemoral vector injection, intravenous administration of LV preferentially targeted short-lived progenitor cells. Myelosuppressive treatment of mice prior to LV-injection did not affect the marking efficiency. Our study represents the first continuous analysis of clonal behavior of genetically marked hematopoietic cells in an unmanipulated system, providing evidence that multiple clones are simultaneously active in murine steady-state hematopoiesis.

Details

Original languageEnglish
Pages (from-to)1961-1970
Number of pages10
JournalStem cells
Volume30
Issue number9
Publication statusPublished - Sept 2012
Peer-reviewedYes
Externally publishedYes

External IDs

PubMed 22696148

Keywords

Keywords

  • Adult stem cells, Hematopoietic stem cells, In vivo marking, Lentiviral vector, Mice, Steady-state hematopoiesis