The purpose of this study was to establish a novel xenotransplant mouse model with human Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL). The bone marrow mononuclear cells (BMMNC) were separated from newly diagnosed Ph(+)ALL patients, and injected into 2.1 Gy of (60)Co irradiated and anti-CD122-conditioned NOD/SCID mice through intra femoral injection. Human hematopoietic chimerism in bone marrow and spleen of the recipients was detected by flow cytometry. Morphological analysis of murine marrow cells were performed using May-Giemsa staining. BCR/ABL1 level was detected by RQ-PCR and FISH assays. Furthermore, leukemia infiltration in the organs was evaluated by hematoxylin-eosin staining, immunohistochemical staining with anti-human CD19 and anti-human CD34 antibodies. The results indicated that the unsorted BMMNC from Ph(+)ALL patients were able to repopulate human Ph(+)ALL in vivo. The percentages of human CD45(+)CD19(+) cells in bone marrow, and spleen of the recipient mice were 87.2% ± 10.1% and 79.9% ± 9.2%, respectively. Furthermore, the engrafted cells possessed same morphology, phenotypic and cytogenetic characteristics as cells from the original Ph(+)ALL patients. Compatible with the clinical features, transplanted Ph(+)ALL cells infiltrated into the brain, liver, and kidney of the recipients. It is concluded that the human-mouse xenotransplant established model using intra femoral injection of an anti-CD122-conditioned NOD/SCID repopulation may provide a promising system to study the biology of human Ph(+)ALL in vivo.