Abstract 4

Characterization of bone marrow stem cell (BMSCs) clones and their potential application for bone tissue engineering

Shobha Mareddy; Yin Xiao*; Ross Crawford

Queensland University of Technology, Brisbane, Australia

The heterogenous nature of BMSCs and their ability to differentiate into multiple mesenchymal lineages generate considerable research interest in the area of stem cell isolation and differentiation after in vitro expansion of BMSCs. Most studies on the molecular characterization of stem cell/ progenitor are based upon the cells derived from colony forming unit fibroblast, which is an assumption that the unit is derived from a single cell. The present study is based on clone culture of stem and progenitor cells obtained from single BMSC cell.

Single cell clone cultures were established in 96 well plates. A total of fourteen fast growing robust clones were selected from three bone marrow samples. It was noted that time taken for 30 population doublings varied for each clone ranging from as less as 35 days to 107 days. Evaluation of their differentiation potential showed 5 clones to be tripotential, 8 appeared to be bipotential and the remaining 1 clone differentiated into only one lineage which could be a committed progenitor. These findings have been further confirmed by the expression of differentiation genes. Flowcytometric analysis showed that the clonal populations have a marker expression that is similar to that of BMSC. Most of them readily expressed CD29, CD44, CD90 and CD105 and are negative for hematopoietic markers like CD34, CD45. This study has identified the most preferred cells that could be used for tissue engineering purposes.

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