In the present study, we successfully established a NOD/SCID mouse model of central nervous system leukemia by injection of acute monocytic leukemia cell line SHI-1 cells into the lateral ventricle. Immunohistochemistry was used to detect human leukocyte common antigen in brain slices. Nested PCR assay was used to detect MLL/AF6 fusion gene expression. After injection, the condition of the mice gradually progressed to cachexia and death (median survival time, 25 days). Leukemic cells were identified in the lung, bone marrow, and lymph node of one mouse. Brain tissue sections showed invasion into the subdural space, pia mater, arachnoid, along the Virchow-Robin space and into the deep brain parenchyma. In summary, a central nervous system leukemia (CNSL) model was established in NOD/SCID mice.