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Figure 3 | Journal of Hematology & Oncology

Figure 3

From: Notch1-induced T cell leukemia can be potentiated by microenvironmental cues in the spleen

Figure 3

Effects of spleen environment on the proliferation and migration of T-ALL cells. (a) T-ALL cells were cultured with normal spleen or BM cells in vitro for the indicated periods, and the GFP+ cells were counted by FACS analysis (n = 5). (b) T-ALL cells were cultured with spleen or BM cells for 24 hours and then isolated and seeded into the upper compartment of a transwell plate, whereas the CM from spleen or BM cells was placed in the lower compartment. The GFP+ cells that migrated into the lower compartment after 24 hours were counted by FACS (n = 5). (c) The expression of migration-related genes in T-ALL cells was measured using real-time PCR after the T-ALL cells were co-cultured with spleen or BM cells for 24 hours. (d) 1 × 108 T-ALL cells were transplanted into non-irradiated mice for 24 hours. The GFP+ cells were isolated from the BM or spleen and the expression of CCR7 and MIP-3β was analyzed by real-time PCR. (e) T-ALL cells were isolated from the BM of splenectomized leukemic mice or the spleen of sham-splenectomized leukemic mice, and an identical number of T-ALL cells was transplanted into two mouse groups (n = 12). The survival curves of the two groups are shown; splenectomy-BM-transplanted and sham-spleen-transplanted groups had a median survival time of 23 and 25.5 days, respectively (p < 0.05). All columns indicate the means ± SD, and the statistical analysis was performed using one-way ANOVA with multiple comparison test, Student’s t test and Kaplan-Meier survival analysis (*, p < 0.05; **, p < 0.01; ***, p < 0.001).

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