Fig. 1.

Cultured BMDC enriched with Tie2⁺/CD45⁺/CD11b⁺cells promote tumor growth. a Experimental design to evaluate the effect of cultured BMDC and fresh bone marrow cells co-inoculated with lung carcinoma cells on tumor growth in immunocompetent mice. b Cultured BMDC promote LAP0297 tumor growth in FVB mice. Mice were inoculated with cancer cells (0.5X10⁶ lung carcinoma LAP0297) alone or either with BMDC (1X10⁶ BMDC +0.5X10⁶ LAP0297) or fresh bone marrow cells (1X10⁶ BM + 0.5X10⁶ LAP0297). Data represent the mean ± SEM; LAP (n = 12), LAP+BMDC (n = 6) and LAP+BM (n = 10); *p < 0.05; ***p < 0.001 (vs LAP); two-way repeated measures ANOVA followed by Tukey test. c BMDCs promote Lewis lung carcinoma (LLC) tumor growth in C57 mice. LLC cells (10⁶) alone or with BMDC (10⁶) were inoculated into immunocompetent C57 mice and tumors allowed to developed for two weeks. Data represent the mean ± SEM of 6 to 7 animals per group. **p < 0.01, ***p < 0.001; LLC vs (LLC + BMDC) analyzed by two-way repeated measures ANOVA followed by Tukey test. d Freshly isolated bone marrow cells attenuate LAP0297 tumor growth. FVB mice were inoculated with LAP0297 cells alone or with freshly isolated bone marrow cells and tumors were monitored for two weeks. Data represent the mean ± SEM of 4 to 5 animals per group. *p < 0.05, ***p < 0.001, two-way repeated measures ANOVA followed by Tukey test. e Rate of green fluorescent cells isolated from Tie2-GFP⁺ transgenic mice. Bone marrow cells from wild type and FVB-Tie2-GFP mice were analyzed by flow cytometry (plots a and b, respectively); plot c shows the rate of fluorescent cells from lungs of FVB-Tie2-GFP transgenic mice. f Cultured BMDC are enriched with Tie2⁺ cells. BMDC and freshly isolated bone marrow cells (BM) from wild type FVB mice were stained with anti Tie2 antibodies and subjected to FACS analysis. g Cultured Tie2⁺/CD45⁺ BMDC express the monocyte marker CD11b. BMDC were stained with antibodies for Tie2 (PE), CD45 (PE-Cy7) and CD11b (FITC) and analyzed by FACS. h Rate of Tie2⁺/CD11b⁺/CD45⁺ cells in LAP0297 tumors. FVB mice were inoculated with 10⁶ LAP0297 cells and tumors allowed to grow for two weeks. Expression of Tie2 (PE), CD45 (PE-Cy7) and CD11b (BV605) in dissected tumors, analyzed as single cell suspensions, was done by FACS. Background fluorescence was assessed with the FITC channel. i Expression of cell markers in BMDCs and freshly isolated bone marrow cells. RT-PCR analysis for the indicated cell markers was performed in fresh bone marrow cells (BM) isolated from FVB mice and in cultured BMDC. Data represent the mean ± SEM of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001; two-way ANOVA followed by the Tukey test