Isolation of bone marrow for a stroke animal model: A step toward autologous transplantation

Mononuclear cells (MNCs) originating in the bone marrow include a population of mesenchymal and hematopoietic stem cells, which secrete a host of cytokines and growth factors involved in natural repair processes. A novel therapeutic intervention that uses an individual's own cells is autologous cell therapy, which has been gaining clinical interest. To autotransplant MNCs in animal models of stroke, the number of MNCs extracted from a femur and the proportion of hematopoietic stem cells to be collected has to be known for determining the optimal number of MNCs that can be extracted from each femur. A total of 10 male, 6-month-old, and 250-400 g Wistar rats were used. We performed a middle cerebral artery occlusion procedure and maintained the rats for one month to isolate MNCs by the Ordodi technique that requires a single femur so that the rat's live is spared. The cells were then processed, counted, and the percentage of CD34⁺ cells determined. The isolation of bone marrow from one rat femur yielded an average 7,190,000 cells, with a proportion of CD34⁺ cells of 1.88%. The survival rate of rats from whom the bone marrow was isolated using this method was 100%. We found in an animal stroke model that autologous MNC transplantation is possible without endangering the rats' lives.