Effect of Bone Marrow Cell Collection Techniques and Donor Site Locations on In-Vitro Growth of Bone Forming Cells

This study was aimed to evaluate the influence of bone marrow cell collection techniques and donor site locations on in-vitro growth of bone forming cells. MATERIAL & METHOD: Sixty six samples of bone marrow cell collections(BMCC) or bone marrow aspirates(BMA) from 15 patients were obtained. Thirty eight samples for culture were composed of 23 BMA from 7 tibial condyles and 16 ilia and other 15 BMCC from contralateral ilia. Other 28 samples were used for the analysis of alkaline phosphatase activities. After counting total cell numbers, mesenchymal stem cells (MSC) obtained from samples were incubated for 14 days. Alkaline phosphatase staining was utilized to count the number of stained colonies. RESULTS: Average MSC counts of BMA from tibial condyles and from ilia and those of BMCC from ilia were 1.42*106, 7.35*106 and 4.80*106 respectively (p=0.010). Culture of MSC aspirated from tibial condyles failed in all 7 samples. However, 9 samples of 15 BMCC and 9 samples of 16 BMA from ilia were successful in culture (p=0.018). The average of cell counts in the successful cultures was 7.92* 106, whereas that in the failed cultures was 2.85*106 (p=0.000). According to the result of multiple regression analysis, colony count was associated with the patient’s age and total cell numbers, but not with collection method such as BMCC or BMA (p=0.000, R=0.648, beta; age=-0.405, cell number=0.356). Based on calculation of the discriminating formula, the number of cells over 5.25*106 was needed for the successful culture. CONCLUSION: For the successful culture in vitro and for graft, the total number of collected bone forming cells is more important than donor sites or collection methods. In young patients, the graft of bone-marrow-derived osteoprogenitor cells is promising.