A rapid method for obtaining mesenchymal stem cells and platelets from bone marrow aspirate.
Mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) are currently used alone or in combination for therapeutic applications especially for bone repair. We tested whether MSCs can be isolated from bone marrow (BM) aspirate using a commercially available kit commonly used to obtain PRP from peripheral blood (PB). Results revealed that mononuclear cells and platelets from both PB and BM could be efficiently isolated by obtaining a mononuclear and platelet rich fraction (PB-MPRF and BM-MPRF, respectively). Starting with comparable volumes, the number of platelets increased 1.5-fold in BM-MPRF compared to PB-MPRF. The number of clonogenic cells in BM-MPRF samples was significantly higher than whole BM samples as revealed by CFU-F assay (54.92 ± 8.55 CFU-F/1.5 x 10(5) nucleated cells and 32.50 ± 12.43 CFU-F/1.5 x 10(5) nucleated cells, respectively). Cells isolated from BM-MPRF after in vitro expansion fulfilled the definition of MSCs by phenotypic criteria, and differentiated along osteogenic, adipogenic and chondrogenic lineages following induction. Results showed that the kit isolated MSCs and platelets from BM aspirate. Isolated MSCs were further expanded in a laboratory and BM-MPRF was used clinically following BM withdrawal for rapid intra-operative cell therapy for the treatment of bone defects.
Bone Regeneration Laboratory, 2nd Clinic of Orthopaedics and Traumatology, Rizzoli Orthopaedic Institute, Bologna, Italy.
Barbara
B
Dozza
Giuliana
G
Gobbi
Enrico
E
Lucarelli
Michela
M
Pierini
Claudia
C
Di Bella
Tommaso
T
Frisoni
Pier Luigi
PL
Tazzari
Francesca
F
Ricci
Prisco
P
Mirandola
Cecilia
C
Carubbi
Sandro
S
Giannini
Davide
D
Donati
Marco
M
Vitale
D000293:Adolescent; D015703:Antigens, CD / Q000378:metabolism; D001707:Biopsy, Needle; D001792:Blood Platelets / Q000166:cytology*; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002648:Child; D020219:Chondrogenesis; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010012:Osteogenesis; D010976:Platelet Count; D055815:Young Adult
D015703:Antigens, CD