cell culture mouse 4t1 mammary tumor cells  (ATCC)

Journal:

Article Title: Massive T-Lymphocyte Infiltration into the Host Stroma Is Essential for Fibroblast Growth Factor-2-Promoted Growth and Metastasis of Mammary Tumors via Neovascular Stability

doi: 10.2353/ajpath.2009.080471

Figure Lengend Snippet: FGF-2 indirectly enhances 4T1 mammary tumor growth and spontaneous metastasis in normal mice but not in nude mice. 4T1 cells were cultured under the serum-starved conditions for 24 hours and then treated with FGF-2. A: Whole cell lysates were collected and immunoblotted with antibodies against FGF receptor 1, phospho-Akt, Akt, phospho-p38, and p38. B: The number of living 4T1 cells was counted at 24 hours after the FGF-2 treatment. Each value represents the mean ± SEM for five samples. Normal and nude female mice (6 weeks old) were inoculated with 4T1 cells into the mammary fat pad, and then FGF-2 was administered once daily at the inoculation site from days 1 to 2 (C) and from days 9 to 10 (D) after the inoculation. Tumor volumes until day 20 and the number of pulmonary metastases on day 30 were evaluated. The timing of FGF-2 administration is represented with a red arrow in each graph of tumor growth. A similar result was obtained in an independent experiment. The number of mice examined is shown in each column. Each value represents the mean ± SEM, and significant differences from each control group are shown as *P < 0.05; **P < 0.01; and N.S, not significant.

Article Snippet: Cell Culture 4T1 mouse mammary tumor cells (4T1 cells) 18 with highly metastatic potentials were obtained from American Type Culture Collection (Manassas, VA), cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 2 mmol/L of l -glutamine, 50 mmol/L of HEPES, 100 U/ml of penicillin, and 100 μg/ml of streptomycin.

Techniques: Cell Culture, Control

Journal:

Article Title: Massive T-Lymphocyte Infiltration into the Host Stroma Is Essential for Fibroblast Growth Factor-2-Promoted Growth and Metastasis of Mammary Tumors via Neovascular Stability

doi: 10.2353/ajpath.2009.080471

Figure Lengend Snippet: FGF-2 induces intensive T-lymphocyte and macrophage infiltration into the host stroma of normal mice. Normal and nude mice were inoculated with 4T1 cells into the mammary fat pad, and then FGF-2 was administered from days 1 to 2 after inoculation using the same procedure described in Figure 1. A: Representative photographs show the immunostaining of CD3-positive T lymphocytes in the host stroma on days 5 and 7. B: The number of T lymphocytes was counted in the host stroma on days 5 and 7. C: Representative photographs show the immunostaining of F4/80-positive macrophages in the host stroma on day 7. D: The number of macrophages was counted in the host stroma on day 7. Five mice were examined in each group. Each value represents the mean number of cells per area (mm2) ± SEM. Significant differences from each control group are shown as *P < 0.05; **P < 0.01; and N.S, not significant. T and S in the photographs represent the tumor tissue and host stroma, respectively.

Article Snippet: Cell Culture 4T1 mouse mammary tumor cells (4T1 cells) 18 with highly metastatic potentials were obtained from American Type Culture Collection (Manassas, VA), cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 2 mmol/L of l -glutamine, 50 mmol/L of HEPES, 100 U/ml of penicillin, and 100 μg/ml of streptomycin.

Techniques: Immunostaining, Control

Journal:

Article Title: Massive T-Lymphocyte Infiltration into the Host Stroma Is Essential for Fibroblast Growth Factor-2-Promoted Growth and Metastasis of Mammary Tumors via Neovascular Stability

doi: 10.2353/ajpath.2009.080471

Figure Lengend Snippet: FGF-2 increases microvessel formation in tumor tissue and the host stroma of normal mice but not nude mice. Normal and nude mice were inoculated with 4T1 cells, and then FGF-2 was administered at the inoculation site from days 1 to 2. A: Representative photographs show the immunostaining of the PECAM-1-positive microvessels developed in the tumor tissue on day 7. B: MVD in the tumor tissue was evaluated on day 7. Each value represents the mean number of cells per area (mm2) ± SEM. Significant differences from each control group are shown as **P < 0.01. C: Representative photographs show the PECAM-1 (e--h) and α-SMA-positive pericyte/VMC (i--l) in the host stroma surrounding the tumor on day 7. The square drawn with the orange lines in the photograph of H&E staining (a--d) show the PECAM-1 and α-SMA photograph area in the host stroma. T and S in the photographs represent the tumor tissue and host stroma, respectively.

Article Snippet: Cell Culture 4T1 mouse mammary tumor cells (4T1 cells) 18 with highly metastatic potentials were obtained from American Type Culture Collection (Manassas, VA), cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 2 mmol/L of l -glutamine, 50 mmol/L of HEPES, 100 U/ml of penicillin, and 100 μg/ml of streptomycin.

Techniques: Immunostaining, Control, Staining

Journal:

Article Title: Massive T-Lymphocyte Infiltration into the Host Stroma Is Essential for Fibroblast Growth Factor-2-Promoted Growth and Metastasis of Mammary Tumors via Neovascular Stability

doi: 10.2353/ajpath.2009.080471

Figure Lengend Snippet: FGF-2-induced COX-2 expression in the host stroma is different between normal and nude mice. Normal and nude female mice were inoculated with 4T1 cells, and then FGF-2 was treated from days 1 to 2. A: The photographs show the immunostaining of COX-2 in the host stroma on days 3 and 5. The square drawn with the orange lines in the photograph of H&E staining on day 3 shows the photograph area of the host stroma-stained COX-2. The T and S represent the tumor tissue and host stroma, respectively. B: The level of COX-2 mRNA in the host stroma was measured on day 3. Each value represents the mean ± SEM for five samples. Significant differences between groups are shown as *P < 0.05 and N.S; not significant.

Article Snippet: Cell Culture 4T1 mouse mammary tumor cells (4T1 cells) 18 with highly metastatic potentials were obtained from American Type Culture Collection (Manassas, VA), cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 2 mmol/L of l -glutamine, 50 mmol/L of HEPES, 100 U/ml of penicillin, and 100 μg/ml of streptomycin.

Techniques: Expressing, Immunostaining, Staining

Journal:

Article Title: Massive T-Lymphocyte Infiltration into the Host Stroma Is Essential for Fibroblast Growth Factor-2-Promoted Growth and Metastasis of Mammary Tumors via Neovascular Stability

doi: 10.2353/ajpath.2009.080471

Figure Lengend Snippet: FGF-2-induced VEGFA expression in the host stroma is different between normal and nude mice. Normal and nude female mice were inoculated with 4T1 cells, and then FGF-2 was administered from days 1 to 2. A: The photographs show the immunostaining of VEGFA in the host stroma on day 5. The T and S represent the tumor tissue and host stroma, respectively. B: The VEGFA mRNA levels in the host stroma were measured on day 5. Each value represents the mean ± SEM for five samples. Significant differences between groups are shown as *P < 0.05 and N.S; not significant. T and S in the photographs represent the tumor tissue and host stroma, respectively.

Article Snippet: Cell Culture 4T1 mouse mammary tumor cells (4T1 cells) 18 with highly metastatic potentials were obtained from American Type Culture Collection (Manassas, VA), cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 2 mmol/L of l -glutamine, 50 mmol/L of HEPES, 100 U/ml of penicillin, and 100 μg/ml of streptomycin.

Techniques: Expressing, Immunostaining

Journal:

Article Title: Massive T-Lymphocyte Infiltration into the Host Stroma Is Essential for Fibroblast Growth Factor-2-Promoted Growth and Metastasis of Mammary Tumors via Neovascular Stability

doi: 10.2353/ajpath.2009.080471

Figure Lengend Snippet: FK506 inhibits FGF-2-induced tumor growth and COX-2 and VEGFA mRNA expression in the host stroma. A: The 4T1 cells were cultured in DMEM, then FK506 (1 and 10 μmol/L) was added to the medium, and the number of cells was counted after 24 hours. B: Mice were inoculated with 4T1 cells in the mammary fat pad. FGF-2 was administered at the inoculation site from days 1 to 2. Tumor volumes (B) until day 20 and the number of pulmonary metastases (C) on day 30 were evaluated. Ten mice were examined in each group. A similar result was obtained in an independent experiment. D: The relative expression level of COX-2 and VEGFA mRNA in the host stroma was measured on day 5 using real-time PCR. Five mice were examined in each group. Each value represents the mean ± SEM. Significant differences between groups are shown as *P < 0.05 and **P < 0.01.

Article Snippet: Cell Culture 4T1 mouse mammary tumor cells (4T1 cells) 18 with highly metastatic potentials were obtained from American Type Culture Collection (Manassas, VA), cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 2 mmol/L of l -glutamine, 50 mmol/L of HEPES, 100 U/ml of penicillin, and 100 μg/ml of streptomycin.

Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction

Journal:

Article Title: Massive T-Lymphocyte Infiltration into the Host Stroma Is Essential for Fibroblast Growth Factor-2-Promoted Growth and Metastasis of Mammary Tumors via Neovascular Stability

doi: 10.2353/ajpath.2009.080471

Figure Lengend Snippet: NS-398 inhibits FGF-2-induced tumor growth, pulmonary metastasis, and VEGFA mRNA expression in the host stroma. A: The 4T1 cells were cultured in DMEM, then NS-398 (2 and 20 μmol/L) was added to the medium, and the number of cells was counted after 24 hours. Mice were inoculated with 4T1 cells in the mammary fat pad. FGF-2 was administered at the inoculation site from days 1 to 2 after inoculation. NS-398 (20 mg/kg) was administered intraperitoneally from days 1 to 9. Tumor volume (B) until day 20 and the number of pulmonary metastases (C) on day 30 were evaluated. Fifteen mice were examined in each group. A similar result was obtained in an independent experiment. D: The relative level of VEGFA mRNA in the host stroma was measured on day 5 using real-time PCR. Five mice were examined in each group. Each value represents the mean ± SEM. Significant differences between groups are shown as **P < 0.01 and *P < 0.05.

Article Snippet: Cell Culture 4T1 mouse mammary tumor cells (4T1 cells) 18 with highly metastatic potentials were obtained from American Type Culture Collection (Manassas, VA), cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 2 mmol/L of l -glutamine, 50 mmol/L of HEPES, 100 U/ml of penicillin, and 100 μg/ml of streptomycin.

Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction