Murine marrow-derived mesenchymal stem cell: isolation, in vitro expansion, and characterization.
In spite of the attention given to the study of mesenchymal stem cells (MSCs) derived from the bone marrow (BM) of humans and other species, there is a lack of information about murine MSCs. We describe the establishment of conditions for the in vitro expansion of plastic-adherent cells from murine BM for over 50 passages, and provide their characterization regarding morphology, surface marker profile and growth kinetics. These cells were shown to differentiate along osteogenic and adipogenic pathways, and to support the growth and differentiation of haematopoietic stem cells, and were thus operationally defined as murine mesenchymal stem cells (mMSCs). mMSCs were positive for the surface markers CD44, CD49e, CD29 and Sca-1, and exhibited a homogeneous, distinctive morphology. Their frequency in the BM of adult BALB/c and C57Bl/6 mice, normal or knockout for the alpha-L-iduronidase (IDUA) gene, was preliminarily estimated to be 1 per 11,300-27,000 nucleated cells. The emergence of a defined methodology for the culture of mMSCs, as well as a comprehensive understanding of their biology, will make the development of cellular and genetic therapy protocols in murine models possible, and provide new perspectives in the field of adult stem cells research.
Meirelles
Lindolfo da Silva
Lda S
Laboratório de Imunogenética, Departamento de Genética, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, Porto Alegre RS, Brasil.
Nardi
Nance Beyer
NB
D000818:Animals; D000950:Antigens, Ly / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology; D002454:Cell Differentiation; D002455:Cell Division; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D018960:Hyaluronan Receptors; D039422:Integrin alpha3 / Q000032:analysis; D019012:Integrin beta1 / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout
D000950:Antigens, Ly; D018960:Hyaluronan Receptors; D039422:Integrin alpha3; D019012:Integrin beta1; C074650:Ly6a protein, mouse; D008565:Membrane Proteins