Human Bone Marrow-derived Mesenchymal Stem Cell: A Source for Cell-Based Therapy

M Ayatollahi, B Geramizadeh, M Zakerinia, M Ramzi, R Yaghobi, P Hadadi, A R Rezvani, M Aghdai, N Azarpira, H Karimi
International Journal of Organ Transplantation Medicine 2012, 3 (1): 32-41

BACKGROUND: The ability of mesenchymal stem cells (MSCs) to differentiate into many cell types, and modulate immune responses, makes them an attractive therapeutic tool for cell transplantation and tissue engineering.

OBJECTIVE: This project was designed for isolation, culture, and characterization of human marrow-derived MSCs based on the immunophenotypic markers and the differentiation potential.

METHODS: Bone marrow of healthy donors was aspirated from the iliac crest. Mononuclear cells were layered over the Ficoll-Paque density-gradient and plated in tissue cultures dish. The adherent cells expanded rapidly and maintained with periodic passages until a relatively homogeneous population was established. The identification of adherent cells and the immune-surface markers was performed by flow cytometric analysis at the third passage. The in vitro differentiation of MSCs into osteoblast and adipocytes was also achieved.

RESULTS: The MSCs were CD11b (CR3), CD45, CD34, CD31 (PCAM-1), CD40, CD80 (B7-1), and HLA-class II negative because antigen expression was less than 5%, while they showed a high expression of CD90, and CD73. The differentiation of osteoblasts, is determined by deposition of a mineralized extracellular matrix in the culture plates that can be detected with Alizarin Red. Adipocytes were easily identified by their morphology and staining with Oil Red.

CONCLUSION: MSCs can be isolated and expanded from most healthy donors, providing for a source of cell-based therapy.

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