title: Mesenchymal content of fresh bone marrow: a proposed quality control method for cell therapy.
abstract: The scarcity of mesenchymal stem cells (MSC) in bone marrow (BM) has justified their ex vivo expansion before therapeutic use, but a method to evaluate the quality of initial mesenchymal content and track the modifications induced by graft processing has not yet been proposed. The aim of this study was to establish such a procedure. Flow cytometric and functional assay methods were modified to count CD45(-) CD14(-)/CD73(+) subsets containing all MSC and used them to study BM from spongy bone (SB) and iliac crest aspirate (ICA). These methods detected the target subsets in all BM suspensions derived from SB (n = 154) and ICA, (n = 44) with a satisfactory correlation between immuno-phenotyping and functional tests by low-density plating. We noted a higher overall MSC frequency in SB cell suspensions but a lower plating efficiency of CD45(-) CD14(-)/CD73(+) SB cells under standard culture conditions. We propose a cell quality control on un-manipulated BM cell suspensions to quantify the mesenchymal compartment with regard to varying donor factors, such as age and sampling site, that influence expansion and define a therapeutic threshold value. Furthermore, we were able to confirm differences in plating efficiency and proliferative capacity between two BM origins.
authors: Hématology (Biology) Department, U.F. de Biologie et Caractérisation Cellulaires, Hotel-Dieu, CHU, Clermont-Ferrand, France.
authors: Richard
authors: R
authors: Veyrat-Masson
authors: Nathalie
authors: N
authors: Boiret-Dupré
authors: Chantal
authors: C
authors: Rapatel
authors: Stéphane
authors: S
authors: Descamps
authors: Laurent
authors: L
authors: Guillouard
authors: Jean-Jacques
authors: JJ
authors: Guérin
authors: Pascale
authors: P
authors: Pigeon
authors: Stéphane
authors: S
authors: Boisgard
authors: Jacques
authors: J
authors: Chassagne
authors: Marc G
authors: MG
authors: Berger
mesh_terms: D015720:5'-Nucleotidase / Q000032:analysis; D001854:Bone Marrow Cells / Q000276:immunology; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D006801:Humans; D007085:Ilium; D017493:Leukocyte Common Antigens; D018950:Lipopolysaccharide Receptors; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000276:immunology; D011786:Quality Control; D033581:Stem Cell Transplantation / Q000592:standards
chemical_list: D018950:Lipopolysaccharide Receptors; D017493:Leukocyte Common Antigens; D015720:5'-Nucleotidase