Culture conditions allow selection of different mesenchymal progenitors from adult mouse bone marrow.
The use of adult stem cells in tissue engineering approaches will benefit from the establishment of culture conditions that allow the expansion and maintenance of cells with stem cell-like activity and high differentiation potential. In the field of adult stem cells, bone marrow stromal cells (BMSCs) are promising candidates. In the present study, we define, for the first time, conditions for optimizing the yields of cultures enriched for specific progenitors of bone marrow. Using four distinct culture conditions, supernatants from culture of bone fragments, marrow stroma cell line MS-5, embryonic fibroblast cell line NIH3T3, and a cocktail of epidermal growth factor (EGF) and platelet-derived growth factor (PDGF), we isolated four different sub-populations of murine BMSCs (mBMSCs). These cells express a well-known marker of undifferentiated embryonic stem cells (Nanog) and show interesting features in immunophenotype, self-renewal ability, and differentiation potency. In particular, using NIH3T3 conditioned medium, we obtained cells that showed impairment in osteogenic and chondrogenic differentiation while retaining high adipogenic potential during passages. Our results indicate that the choice of the medium used for isolation and expansion of mBMSCs is important for enriching the culture of desired specific progenitors.
Esposito
Maria Teresa
MT
CEINGE Biotecnologie Avanzate sca r l, Naples, Italy.
Di Noto
Rosa
R
Mirabelli
Peppino
P
Gorrese
Marisa
M
Parisi
Silvia
S
Montanaro
Donatella
D
Del Vecchio
Luigi
L
Pastore
Lucio
L
D050156:Adipogenesis / Q000187:drug effects; D000375:Aging / Q000187:drug effects / Q000502:physiology*; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D018398:Homeodomain Proteins / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D041681:NIH 3T3 Cells; D000071317:Nanog Homeobox Protein; D010012:Osteogenesis / Q000187:drug effects
D017077:Culture Media, Conditioned; D018398:Homeodomain Proteins; D053605:Leukemia Inhibitory Factor; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse