Study of the osteogenic capacity of human normal bone marrow mesenchymal stroma cells.

HOFER EL.; CARAMUTTI D.; LARUSSA V.; FELDMAN L.; CHASSEING NA.

Book of Stem Cells: Research tools and transplantation. A serie from Methods in Molecular Biology Books.

The Humana Press.

Lugar: Totowa, NJ07512, USA; Año: 2007;

Abstract Bone Marrow (BM) - mesenchymal stromal cells (MSC= mesenchymal stem cells and mesenchymal stromal progenitors) play an important role on bone formation as being the major source of osteoprogenitor cells. In the present paper we described the detail practical procedure to study the osteogenic potential of normal human BM-MSC. MSC derived from low density 3rd passage were cultured for 21 days in osteogenic medium. Osteogenesis was analyzed by Von Kossa and Giemsa stains. The parameters evaluated were: colonies with osteogenic cells, % of Petri dish surface with differentiated cells and pattern of differentiation (grades: 0= fibroblasts, 1= cell in the way to osteoblasts, 2= osteoblasts and 3= osteocytes). 100% of normal volunteers MSC cultures developed colonies with osteogenic cells and showed a 24±4% of the Petri dish surface with differentiated cells. Moreover, the 43% of normal cultures reached grade 1 and the other 57% grades 2-3. The osteogenic cultures developed calcium deposits and the mineralization matrix with a homogeneous distribution over the complete surface of the Petri dish. Key words: mesenchymal stromal cells, bone marrow, osteoblast, osteocyte.