High recovery of mesenchymal progenitor cells with non-density gradient separation of human bone marrow.
OBJECTIVE
Bone marrow (BM) is the most used source of hemopoietic stem cells (HSC) and mesenchymal stromal cells (MSC) in both hematologic settings and regenerative medicine. We compared the feasibility and reproducibility of two gravity separation techniques, with or without the use of a density gradient, in terms of both hematopoietic and mesenchymal human BM progenitors.


METHODS
A total of 16 BM samples was processed to obtain mononuclear cells (MNC) and buffy coats (BC). The efficiency of the two procedures was evaluated by recovery of white blood cells (WBC), MNC and CD34(+) cells, clonogenic assays, red blood cell (RBC) depletion, cell viability, expression of embryonic transcriptional regulators and MSC assessment.


RESULTS
The two procedures yielded a comparable recovery of HSC. Non-density gradient separation (NDGS) of BM resulted in four times higher MSC recovery and higher expression of embryonic stem cell markers (Nanog and Sox2) compared with density-gradient separation (DGS). MSC derived from both procedures was comparable in terms of phenotype, differentiation and proliferation potential.


CONCLUSIONS
NDGS is less time consuming, provides a better MSC enrichment and appears to be a suitable cell preparation method for clinical applications.
Dal Pozzo
Simone
S
Department of Hematology, Careggi Hospital, Florence, Italy.
Urbani
Serena
S
Mazzanti
Benedetta
B
Luciani
Paola
P
Deledda
Cristiana
C
Lombardini
Letizia
L
Benvenuti
Susanna
S
Peri
Alessandro
A
Bosi
Alberto
A
Saccardi
Riccardo
R
D015415:Biomarkers / Q000378:metabolism; D001853:Bone Marrow / Q000473:pathology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D005240:Feasibility Studies; D005434:Flow Cytometry; D006112:Gravitation; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000378:metabolism / Q000473:pathology*; D015203:Reproducibility of Results
D015415:Biomarkers