normal lung fibroblast cells wi38  (ATCC)

Journal: eLife

Article Title: A subset of CB002 xanthine analogs bypass p53-signaling to restore a p53 transcriptome and target an S-phase cell cycle checkpoint in tumors with mutated-p53

doi: 10.7554/eLife.70429

Figure Lengend Snippet: HCT116 isogenic panel treated with CB002 or analog #4 for 48 hr and their respective IC50 values shown in the table ( A ). CB002-analog #4 increases apoptotic cells as indicated by the sub-G1 content in cancer cells but not in normal WI38 cells (48 hr). Two-way ANOVA, p<0.0001 ( B ). 72 hr treatment with CB002-analog #4 is most potent ( C ) and increases dead cells as indicated by the ethidium homodimer staining (red) compared to calcein stained live cells (green) ( A ), and cleaved caspase-3 (green) immunofluorescence ( D ) in colorectal cancer patient-derived organoid cells. CB002-analog #4 decreases ki67 staining (green) in a dose-dependent manner (72 hr) in colorectal cancer patient-derived organoid cells ( E ). CB002-analog #4 is non-toxic in vivo ( F ) and significantly reduces tumor volume in NSG mouse xenografts with SW480 wild-type cells ( G ) but not in SW480 cells with shNoxa ( H ). 50 mg/kg by oral gavage three times per week, final tumor volume at 5 weeks. Unpaired t-test, p<0.05.

Article Snippet: DLD-1 (p53 S241F ) (RRID: CVCL_0248 ), SW480 (p53 R273H,P309S ) (RRID: CVCL_0546 ), and HCT116 (p53 WT ) (RRID: CVCL_0291 ) colorectal cancer cell lines and WI38 normal lung fibroblast cells were purchased from ATCC.

Techniques: Staining, Immunofluorescence, Derivative Assay, In Vivo

Journal: Scientific reports

Article Title: Primary lung cancer samples cultured under microenvironment-mimetic conditions enrich for mesenchymal stem-like cells that promote metastasis.

doi: 10.1038/s41598-019-40519-4

Figure Lengend Snippet: Figure 1. Physiological hypoxia and cell derived matrix enhance proliferation of stromal cell lines and are non- desmoplastic. (A) Schematic of cell derived matrix preparation from IMR90 fetal lung fibroblast cell line and characteristics of microenvironment mimetic conditions. (B) Phase contrast images of equal numbers WI38 fetal lung fibroblast cells grown for 72 hours in four environmental conditions (2D Normoxia, 2D Hypoxia, 3D Normoxia, and 3D Hypoxia). Scale bars are 100 μm. (C) Doubling time of WI38, IMR90, and bone marrow mesenchymal stem cell line hMSC following short term growth in different environments (Statistical analysis performed by ANOVA with Dunnett’s test for multiple comparison to 2DN, *p < 0.05, **p < 0.01, and ****p < 0.0001; data presented as mean ± S.E.M.). (D) Western blot analysis of hypoxia inducible factor 1 alpha (Hif1a), alpha smooth muscle actin (α-SMA), and cell cycle inhibitors (p21, p27) from WI38 and IMR90 after 72 hours growth in different environments. Cropped images in the figure are derived from full length blots presented in Supplementary Fig. S3.

Article Snippet: Human normal fetal lung fibroblast cell lines WI38 and IMR90 were purchased from American Type Culture Collection (ATCC) and maintained in Minimum Essential Medium Alpha modification (MEM-α) supplemented with L-Gln, ribo- and deoxyribonucleosides (Fisher), 10% fetal bovine serum (Invitrogen) and 1% antibiotic/antimycotic (Sigma).

Techniques: Derivative Assay, Comparison, Western Blot

Journal: Scientific reports

Article Title: Primary lung cancer samples cultured under microenvironment-mimetic conditions enrich for mesenchymal stem-like cells that promote metastasis.

doi: 10.1038/s41598-019-40519-4

Figure Lengend Snippet: Figure 6. Cell-cell interactions between 3DH Patient derived stromal cell lines and lung adenocarcinoma promote cancer cell proliferation and colony formation. Stromal cells were grown to a confluent monolayer for four days, and then equal amounts of red nuclear labeled A549 cells were plated atop monolayers. (A) (Upper) Phase contrast and fluorescent images of A549 grown on stromal monolayers of IMR90 and patient stroma 959. Images obtained 1 and 7 days after cancer cell seeding. (Lower) Comparison of A549 colony formation on IMR90 or 955 monolayers on day 7. (B) Comparison of A549 colony formation on hMSC, 956, and 960 monolayers. (C) Quantification of red nuclei on day 7 (mean nuclear counts were obtained from multiple images of technical duplicate plates from 2 different experiments.) Statistical analysis performed by ANOVA with Tukey’s test for multiple comparisons to the indicated groups (a: vs. IMR90, P < 0.01; b: vs WI38, P < 0.05; c: vs. hMSC, P < 0.05, d: vs. 959, P < 0.01); data presented are mean ± S.E.M. All scale bars are 100 μm.

Article Snippet: Human normal fetal lung fibroblast cell lines WI38 and IMR90 were purchased from American Type Culture Collection (ATCC) and maintained in Minimum Essential Medium Alpha modification (MEM-α) supplemented with L-Gln, ribo- and deoxyribonucleosides (Fisher), 10% fetal bovine serum (Invitrogen) and 1% antibiotic/antimycotic (Sigma).

Techniques: Derivative Assay, Labeling, Comparison