Characterization of nonexpanded mesenchymal progenitor cells from normal adult human bone marrow.
OBJECTIVE
Adult bone marrow (BM) mesenchymal stem/progenitor cells (MS/PC) are a potentially useful tool for cell therapy and tissue repair. However, the identification of cell subsets rich in MS/PC from fresh BM has not been described. We have developed a means of identifying such subsets from untouched bone marrow.


METHODS
First, MS/PC were enriched by short-time adherence (D(1-3)) before any cell division to evaluate the efficiency of CD73, CD105, CDw90, and CD49a antigens to select highly purified CD45(-)CD14(-) fluorescence-activated sorted subsets enriched in clonogenic mesenchymal cells. Then, we adapted this method to unmanipulated BM mononuclear cells (MNC).


RESULTS
Short-time (D(1-3)) adherent CD45(-)CD14(-) cells expressing CD73 or CD49a antigens contained all the CFU-F, even though the CD105(+) and CDw90(+) subsets comprised less than half the total. In fresh unmanipulated BM MNC, CD73 and CD49a were also highly discriminative and allowed up to a 3 log enrichment of CFU-F when compared to BM MNC. Normal culture conditions upregulated most of the tested antigens.


CONCLUSIONS
The CD45(-)CD14(-)/CD73(+) and CD45(-)CD14(-)/CD49a(+) phenotypes identified subsets containing all the CFU-F and sufficiently enriched to detect them in fresh BM, enabling evaluation of mesenchymal content of BM collections for cell therapy.
Boiret
Nathalie
N
Hématologie Biologique, U.F. de Biologie et Caractérisation Cellulaires, Faculté de Médecine et de Pharmacie, Clermont-Ferrand, France.
Rapatel
Chantal
C
Veyrat-Masson
Richard
R
Guillouard
Laurent
L
Guérin
Jean-Jacques
JJ
Pigeon
Pascale
P
Descamps
Stéphane
S
Boisgard
Stéphane
S
Berger
Marc Gabriel
MG
D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology*; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D016130:Immunophenotyping; D008648:Mesoderm / Q000166:cytology*; D012016:Reference Values
D015703:Antigens, CD