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Identification and isolation from either adult human bone marrow or G-CSF−mobilized peripheral blood of CD34+/CD133+/CXCR4+/ LinCD45 cells, featuring morphological, molecular, a

Author:
Hanna Sovalata, hsovalat@yahoo.fr, Maurice Scrofania, Antoinette Eidenschenka, Sté  phanie Pasqueta, Valé  rie Rimelenb,Philippe Hé  nona  


Journal:
Experimental Hematology


Issue Date:
2011


Abstract(summary):

Objective

Recently, we demonstrated that normal human bone marrow (hBM)−derived CD34+ cells, released into the peripheral blood after granulocyte colony-stimulating factor mobilization, contain cell subpopulations committed along endothelial and cardiac differentiation pathways. These subpopulations could play a key role in the regeneration of post-ischemic myocardial lesion after their direct intracardiac delivery. We hypothesized that these relevant cells might be issued from very small embryonic-like stem cells deposited in the BM during ontogenesis and reside lifelong in the adult BM, and that they could be mobilized into peripheral blood by granulocyte colony-stimulating factor.

Materials and Methods

Samples of normal hBM and leukapheresis products harvested from cancer patients after granulocyte colony-stimulating factor mobilization were analyzed and sorted by multiparameter flow cytometry strategy. Immunofluorescence and reverse transcription quantitative polymerase chain reaction assays were performed to analyze the expression of typical pluripotent stem cells markers.

Results

A population of CD34+/CD133+/CXCR4+/Lin CD45 immature cells was first isolated from the hBM or from leukapheresis products. Among this population, very small (2−5 μm) cells expressing Oct-4, Nanog, and stage-specific embryonic antigen-4 at protein and messenger RNA levels were identified.

Conclusions

Our study supports the hypothesis that very small embryonic-like stem cells constitute a “mobile” pool of primitive/pluripotent stem cells that could be released from the BM into the peripheral blood under the influence of various physiological or pathological stimuli. In order to fully support that hBM- and leukapheresis product−derived very small embryonic-like stem cells are actually pluripotent, we are currently testing their ability to differentiate in vitro into cells from all three germ layers.



Page:
495-505


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