Bone marrow cells in cultures were divided into four groups and cultured with various cytokines in vitro. These four groups are: control, IL-2 group, CD3-AK group, and CIK group. The morphological (cell volume, nucleus/plasm) changes of bone marrow cells in culture were observed. Immunophenotype analysis (CD34, CD38, CD3, CD56) were done before and after culture in all groups. Cytotoxicity against fresh acute leukemia cells were detected by modified MTT methods. The cell volume became larger with increased nucleus/plasm ratio in IL-2 group, CD3-AK group and CIK group. The plasm filled with PAS positive granules in most of cells in CD3-AK group and CIK group. The positive ratio of CD3, CD56, CD38 in CD3-AK or CIK group increased markedly after culture (P < 0.05), but no significant difference between the two groups. The CD56(+) cell increased in IL-2 group. CD34(+) cells decreased in all groups and there were no significant differences among those four groups. The cytotoxicity to fresh leukemia cells: CD3-AK group and CIK group > IL-2 group > control group. There was no significant difference between CD3-AK group and CIK group. This experiment showed different effect on bone marrow cells by different cytokine combination. The cytokine combination of CD3-AK group or CIK group can make immunocytes of bone marrow proliferating and retained certain amount of stem/progenitor cells.