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Bone marrow stromal stem cells lack of specific surface marker and their identification challenges scholars all the time.


To explore the culture condition for clonal isolation of human bone marrow stromal stem cells (BMSCs) and identify their surface markers and differentiation potentials.


Human bone marrow was taken and BMSCs were isolated using density gradient centrifugation. Clone-like cells were selected by single cell limiting dilution. The surface antigens of the cloned BMSCs were analyzed by flow cytometry and immunocytochemistry. Multi-differentiation capacities were evaluated by inducing BMSCs into osteoblasts, chondrocytes and cadiocytes. RT-PCR and immunocytochemistry were applied to detect the three mutli-differentiations.


Single cell-derived BMSCs could be expanded to passage 28 in vitro which still maintaued active proliferation ability. The expanded BMSCs expressed CD29, CD44, CD106, but not CD14, CD34, CD45, HLA-DR. They could be induced to differentiate into osteoblasts, chondrocytes and cadiocytes. These findings suggest that cloned BMSCs can maintain the phenotypes of stem cells during in vitro culture.

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