Investigation of Biological Characteristics and Culture of Stromal Cells of Human Bone Marrow in Vitro

Objective: To establish a model of osteoblasts culture in vitro and investigate biological characteristics of cultured adult bone marrow stromal cells(MSC).That will be experimental basis of MSC used as the functional cells in bone tissue engineering. Methods: Bone marrow was obtained by aspirated in the posterior superior iliac spine or flushed medullary cavity in costal bone from an adult patient whose marrow is normal. Mononuclear cells were then isolated by centrifugation for primary culture in different culture fluid systems. Subculture was done for 8 passages consecutively. During the culture the cells were examined by phase-contrast microscope, tetracycline fluorescence label, histochemical stains. Results: Different kinds of shapes were observed in the cultured cells. Clumps and multilayer of cells became evident and the cell nodules were formed. Alkaline phosphatase staining showed strong positive reaction in cytoplasm. Tetracycline fluorescence label showed positive reaction in the nodules. Conclusions: The morphological and biological characteristics of the cultured cells derived from adult bone marrow were similar to those of osteoblasts, and the cells could synthesize mineralized tissue in vitro.