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Aislamiento y caracterización de células “stem” mesenquimales de médula ósea humana según criterios de la Sociedad Internacional de Terapia Celular

Author(s): Vivian Marcela Rodríguez-Pardo | María Fernanda Fuentes-Lacouture | Jose Alejandro Aristizabal-Castellanos

Journal: Universitas Scientiarum
ISSN 0122-7483

Volume: 15;
Issue: 3;
Start page: 224;
Date: 2010;
Original page

Keywords: stem cells | mesenchymal cells | flow cytometry | bone marrow | cell differentiation

Isolation and characterization of mesenchymal stem cells from human bone marrow according to the criteria of the InternationalSociety for Cellular Therapy. Bone marrow (BM) is an important source for isolating mesenchymal stem cells (MSC) useful inimmunomodulation and tissue regeneration therapies. Objective. To isolate and characterize mesenchymal stem cells obtained from BMmeeting the requirements of the International Society for Cellular Therapy. Materials and methods. BM samples were collected fromvolunteer donors attending the Orthopedics Service of the San Ignacio University Hospital (Bogotá, Colombia). Morphological characteristicswere evaluated by inverted microscopy and the immunophenotype was determined by flow cytometry. Protocols were developed foradipogenic, osteogenic and chondrogenic differentiation using the Oil Red O, alkaline phosphatase and safranin stains, respectively.Results. We collected 24 samples of BM from patients with total hip replacement (volume of BM sample: 5-45 ml). Cells with afibroblastoid morphology were isolated from 21 BM samples (isolation efficiency: 87.5%). No statistical significant differences were foundbetween the hematopoyetic antigens (CD34 and CD45, p>0.05) in the immunophenotypic evaluation (of MSC from BM); on the contrary,there were differences (p=0.006) between the hematopoyetic antigen CD45 and the mesenchymal antigens (CD13, CD44, CD73, CD90,CD105, HLA-I, and HLA-DR). Oil Red O stain revealed the presence of multilocular adipocytes, in the osteogenic induction we observedlocalized mineralization nodules, and chondrogenesis was positive as revealed by the safranin stain. Conclusion. MSC were satisfactorilyisolated from BM and characterized according to the international standards.
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