Author/Authors :
Ayatollahi, M. shiraz university of medical sciences - Transplant Research Center, شيراز, ايران , Geramizadeh, B. shiraz university of medical sciences - Transplant Research Center, شيراز, ايران , Zakerinia, M. shiraz university of medical sciences - Hematology Research Center - Bone Marrow Transplantation Unit, شيراز, ايران , Ramzi, M. shiraz university of medical sciences - Hematology Research Center - Bone Marrow Transplantation Unit, شيراز, ايران , Yaghobi, R. shiraz university of medical sciences - Transplant Research Center, شيراز, ايران , Hadadi, P. shiraz university of medical sciences - Transplant Research Center, شيراز, ايران , Rezvani, A. R. shiraz university of medical sciences - Hematology Research Center - Bone Marrow Transplantation Unit, شيراز, ايران , Aghdai, M. shiraz university of medical sciences - Transplant Research Center, شيراز, ايران , Azarpira, N. shiraz university of medical sciences - Transplant Research Center, شيراز, ايران , Karimi, H. shiraz university of medical sciences - Transplant Research Center, شيراز, ايران
Abstract :
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 lay- ered over the Ficoll-Paque density-gradient and plated in tissue cultures dish. The adherent cells ex- panded 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 adipo- cytes 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 cellbased therapy.
