wi38 fetal lung fibroblasts  (ATCC)

Journal: Aging cell

Article Title: hTERT Increases TRF2 to Induce Telomere Compaction and Extend Cell Replicative Lifespan.

doi: 10.1111/acel.70105

Figure Lengend Snippet: FIGURE 3 | hTERT variants regulate the TRF2:TRF1 ratio by transcriptional and posttranslational mechanisms. (A) TRF2 mRNA levels deter- mined by RT-PCR in young and senescent BJ, Hs68, and Wi38 fibroblast strains. (B) Western blots of young (Y) and senescent (S) fibroblast strains probed with the indicated antibodies. (C) TRF1 mRNA levels in young and senescent BJ cells and BJs expressing WT-hTERT, R865C-hTERT, and V144M-hTERT variants. (D) Western blots of young and senescent BJ fibroblasts, and BJ cells expressing hTERT variants, probed with the indicated antibodies. (E) TRF2 protein levels in young and senescent fibroblast strains. (F) TRF2 protein levels in young and senescent BJs and BJs expressing hTERT variants. (A,C,E,F) Error bars = 95% CI; n = 3. p values determined by two-tailed unpaired t-tests with Welch's correction, ns = not significant. (G) Quantitation of the levels of the CDC20 and (H) FBXO5 Siah1 e3 ubiquitin ligases in cells expressing hTERT variants. Graphs show three bio- logical replicates with tables showing densitometry values for each replicate, with young BJ fibroblasts set to a value of 1. Error bars = 95% CI; n = 3. p values determined by two-tailed unpaired t-tests with Welch's correction. (I) Mock-transfected cells or cells expressing Siah1, CDC20, or FBXO5 were harvested 48 h after transfection, and lysates were blotted for the indicated proteins. GAPDH served as the loading control.

Article Snippet: Primary Hs68 human foreskin fibroblasts (ATCC CRL- 1635) and WI38 fetal lung fibroblasts (ATCC CCL- 75) were grown in DMEM, 1 g/L glucose (Gibco, 11885- 084), supplemented with 10% (v/v) FBS.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Two Tailed Test, Quantitation Assay, Ubiquitin Proteomics, Transfection, Control

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