Abstract:

BACKGROUND: The number of adult bone marrow mesenchymal stem cells (BMSCs) is few. To apply BMSCs to tissue engineering and cell therapy, we should establish mature, simple and effective isolating culture amplification system.
OBJECTIVE: To establish a method of isolation and cultivation in vitro of adult BMSCs, and to observe the results of BMSCs differentiating into the adipocytes, osteoblasts and chondrocytes.
METHODS: Adult BMSCs were respectively isolated and cultivated by density gradient centrifugation and whole bone marrow culture. Cell morphology was observed under an optical microscope to draw growth curves. The effects of different culture methods on BMSCs were compared. CD34, CD44 and CD105 expressions were detected by immunofluorescence. Adult BMSCs were induced with different media. Differentiations of BMSCs into adipocytes, osteoblasts and chondrocytes were measured using Oil Red O staining, alizarin red staining and alcian blue staining.
RESULTS AND CONCLUSION: By the method of whole bone marrow culture, the characteristics of proliferation and growth of BMSCs were superior to that by the method of density gradient centrifugation. Immunophenotypically, this cell population was found to be highly positive for CD44 and CD105, while negative for CD34. Following induced culture, Oil Red O staining, alizarin red staining and alcian blue staining were positive, which verified that BMSCs harvested by the method of whole bone marrow culture gained more capacity of growth and proliferation, and BMSCs could be induced to differentiate into osteoblasts, adipocytes and chondrocytes.