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

PURPOSE: This study evaluated the influence of bone marrow cell collection techniques and donor site locations on the in-vitro growth of bone-forming cells. METHODS: 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, with the other 15 BMCC from the contralateral ilia. The other 28 samples were used for the analysis of alkaline phosphatase activities. After counting total cell number, mesenchymal stem cells (MSC) obtained from samples were incubated for 14 days. Alkaline phosphatase staining was used to count the number of stained colonies to show osteogenic differentiation. RESULTS: The average MSC counts of BMA from tibial condyles and ilia were 1.42×10(6) and 7.35×10(6) respectively, with 4.80×10(6) from ilial BMCC (p=0.010). MSC cultures could not be produced from tibial condyles in all 7 samples. However, 9 of 15 BMCC samples and 9 of 16 ilial BMA samples were successfully cultured (p=0.018). The average of cell counts in the successful cultures was 7.92×10(6), whereas that in the failed cultures was 2.85×10(6) (p=0.000). Multiple regression analysis showed that colony count was associated with the patient's age and total cell numbers, but not with collection methods such as BMCC or BMA (p=0.000, R=0.648, beta; age=-0.405, cell number=0.356). The discriminating formula indicated that more than 5.25×10(6) cells were needed for successful culture. CONCLUSIONS: For successful cultures in vitro and for grafts, the total number of collected bone forming cells is more important than donor sites or collection methods. For young patients, grafting of bone-marrow-derived osteoprogenitor cells is promising.