murine embryonic fibroblasts cell line nih 3t3  (ATCC)

Journal: Molecular Therapy Oncology

Article Title: Collagen depletion by pirfenidone enhances antitumor effect of oncolytic adenovirus against peritoneal metastases of gastric cancer

doi: 10.1016/j.omton.2025.201045

Figure Lengend Snippet: GC cells increase collagen 1 expression in fibroblasts and collagen increases proliferation of GC cells (A) Expressions of Col1A1, Col1A2, and ACTA2 mRNA in mouse gastric cancer (GC) cells (T3-2D), human GC cells (MKN45, MKN7, and NUGC4), mouse fibroblast cells (MEF), and human fibroblast cells (YS-1 and FEF3) are shown. All cells were analyzed by quantitative RT-PCR analysis. Data are expressed as the mean ± SD ( n = 3). (B) Expression of Col1A1 and Col1A2 mRNA in MEF (upper) and YS-1 (lower) cells after incubation with serum-free medium (SFM) as a control and conditioned medium (CM) of each GC cell type for 4 days. Cells were analyzed using quantitative RT-PCR analysis. Data are expressed as the mean ± SD ( n = 3). (C) Representative images of immunocytochemical staining of collagen 1 and α-SMA in MEF (left) and YS-1 (right) after incubation with normal medium and CM of each of the GC cells for 4 days. SFM was used as a control. The area index for each staining was evaluated by ImageJ software. Data are expressed as the mean ± SD ( n = 3). Scale bars, 50 μm. (D) Cell proliferation of T3-2D and MKN45 cells with and without 10% collagen stimulation for 72 h. Data are expressed as the mean ± SD ( n = 5). (E) Representative cellular morphological images of T3-2D and MKN45 cells with and without 10% collagen 1 stimulation for 72 h. Scale bar, 100 μm. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: A murine embryonic fibroblast (MEF) cell line was purchased from the American Type Culture Collection (Manassas, VA) and maintained in DMEM supplemented with 15% FBS.

Techniques: Expressing, Quantitative RT-PCR, Incubation, Control, Staining, Software

Journal: Molecular Therapy Oncology

Article Title: Collagen depletion by pirfenidone enhances antitumor effect of oncolytic adenovirus against peritoneal metastases of gastric cancer

doi: 10.1016/j.omton.2025.201045

Figure Lengend Snippet: Collagens and fibroblasts inhibit oncolytic virus penetration and reduce antitumor effects (A) T3-2D, MKN45, MEF, and YS-1 cells were infected with OBP-702 at the indicated MOIs for 3 days. Cell viability was assessed using the XTT assays. Cell viability was calculated relative to that of the mock-infected cells, which were set as 1.0. Data are expressed as the mean ± SD ( n = 5). (B) T3-2D and MKN45 cells were infected with OBP-702 at the indicated MOIs for 3 days with or without 10% collagen containing medium. Cell viability was assessed using the XTT assays. Cell viability was calculated relative to that of the mock-infected cells, which were set as 1.0. Data are expressed as the mean ± SD ( n = 5). (C) Representative cellular morphological images of T3-2D and MKN45 cells infected with 100 MOI of OBP-702 with or without 10% collagen stimulation for 72 h. Scale bars, 100 μm. (D) Representative spheroid morphological images of T3-2D cells infected with 20 MOI of OBP-702 or MKN45 cells infected with 50 MOI of OBP-702 with or without 2% collagen stimulation for 7 days. Scale bars, 100 μm. (E) ATP cell viability assay of T3-2D and MKN45 spheroids with or without 2% collagen stimulation for 7 days after infection with OBP-702 (20 MOI or 50 MOI). Data are expressed as the mean ± SD ( n = 5). (F) Representative images of mono-spheroids and co-cultured spheroids infected with OBP-401 (100 MOI) for 48 h. T3-2D and MKN45 cells were labeled with red cell trackers and MEF and YS-1 cells with blue cell trackers. Scale bars, 50 μm. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: A murine embryonic fibroblast (MEF) cell line was purchased from the American Type Culture Collection (Manassas, VA) and maintained in DMEM supplemented with 15% FBS.

Techniques: Virus, Infection, Viability Assay, Cell Culture, Labeling