Journal: Frontiers in Medicine

Article Title: Stromal cells support the survival of human primary chronic lymphocytic leukemia (CLL) cells through Lyn-driven extracellular vesicles

doi: 10.3389/fmed.2022.1059028

Figure Lengend Snippet: Influence of tyrosine kinase inhibitor Dasatinib on the release of extracellular vesicles (EVs) in stromal cells. HS-5 (A) and WI-38 (B) cells (2 × 10 6 /ml) were cultivated at 37°C in serum-free RPMI-1640 with dilution series of Dasatinib as indicated. DMSO (0.1% in PBS) served as a control. After 24 h, cells and the supernatant were separated by centrifugation (10 min, 300 × g ). The supernatant was cleared by centrifugation at 3,000 × g and afterwards subjected to ultracentrifugation (90 min, 110,000 × g ). After ultracentrifugation, the EV pellet was suspended in PBS and the count was determined by nanoparticle tracking analysis. The cell pellet was used for cell viability determination. The viability of the DMSO-treated aliquots served as controls (100%). The statistic evaluation was performed by the Kruskal–Wallis test.

Article Snippet: The cell lines StromaNKtert (CVCL_4667), HS-5 (CRL-11882), WI-38 (ATCC#CCL-75), MEC-1 (CVCL_1870) were cultivated at 37°C and 5% CO 2 in RPMI 1640 containing 10% FBS, supplemented with GlutaMAX (2 mM), 100 U/ml penicillin and 100 μg/ml streptomycin.

Techniques: Centrifugation

Journal: Scientific Reports

Article Title: Cellular uptake and retention studies of silica nanoparticles utilizing senescent fibroblasts

doi: 10.1038/s41598-022-26979-1

Figure Lengend Snippet: Cellular uptake kinetics of SiNPs of the 200 nm size range in proliferative and senescent WI-38 fibroblasts over a 72 h timecourse. ( a ) Electron micrographs of SiNPs. The histogram to the right shows TEM and DLS measurements. ( b ) Staining for SA-β-gal activity at 48 h to confirm cellular senescence (bluish-green stain). ( c ) Flow cytometry results for cells incubated continuously with 6.25 µg/ml of SiNPs. Error bars indicate S.E.M. (n = 3). ( d ) Box plot for cell size indexes over time (n = 25 for each condition) . ( e ) Graph shows the amount of cell proliferation over time. Error bars indicate S.E.M. (n = 3).

Article Snippet: We obtained the WI-38 (CCL-75) cell line from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Staining, Activity Assay, Flow Cytometry, Incubation

Journal: Scientific Reports

Article Title: Cellular uptake and retention studies of silica nanoparticles utilizing senescent fibroblasts

doi: 10.1038/s41598-022-26979-1

Figure Lengend Snippet: SiNP retention inside senescent cells and transfer between senescent cells is negligible. ( a ) Proliferative WI-38 fibroblasts were incubated with 25 μg/ml SiNPs for 80 min. On day 1, the cells with SiNPs (+SiNPs) were analyzed by flow cytometry alongside control cells with no silica NPs (−SiNPs). ( b ) The +SiNPs cells from ( a ) were split into two treatment protocols for continued growth and senescence induction (see “ ” section for details). After 8 days in culture, flow cytometry revealed senescent cells had retained their SiNPs while proliferative cells diluted their content with each cell division. ( c ) Representative flow cytometry histogram showing the fluorescence intensity of +SiNPs senescent cells after 10 days compared to −SiNPs control senescent cells without. The +SiNPs and −SiNPs populations were mixed and analyzed by flow cytometry at 0 h and 24 h. The +SiNP population between 0 to 24 h increased by 8.6 ± 4.4% (n = 3).

Article Snippet: We obtained the WI-38 (CCL-75) cell line from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Incubation, Flow Cytometry, Fluorescence

Journal: Virology Journal

Article Title: Isolation and genetic characterization of human coronavirus NL63 in primary human renal proximal tubular epithelial cells obtained from a commercial supplier, and confirmation of its replication in two different types of human primary kidney cells

doi: 10.1186/1743-422X-10-213

Figure Lengend Snippet: Appearance of early RPTEC culture and of WI-38 cells during bioactive agent release assay. [ A ] Vacuolated RPTEC cells, 24 hr culture, 400X. [ B ] Non-infected WI-38 cells demonstrating expected fibroblast shapes, 400X. [ C ] WI-38 cells 12 hrs post-exposure to spent BGM from a 24 hr RPTEC culture, 400X.

Article Snippet: Cell lines A549 (CCL-185), BHK-21 (CCL-10), CaCo-2 (HTB-37), CV-1 (CCL-70), HEK-293 (CRL-1573), LLC-MK2 (CCL-7), MDCK, (CCL-34), Mv1 Lu (CCL-64), NIH/3 T3 (CRL-1658), Vero E6 (CRL-1586), and WI-38 (CCL-75) were obtained from the ATCC (Manassas, VA), and along with MDCK-London cells, were propagated as monolayers at 37°C and 5% CO 2 in Dulbecco's Modified Eagle's Medium (DMEM) (Mediatech, Inc., Manassas, VA) or Eagle’s Minimal Essential Medium (EMEM) (Invitrogen Corp., Carlsbad, CA), as appropriate per cell line.

Techniques: Release Assay, Infection

Journal: Virology Journal

Article Title: Isolation and genetic characterization of human coronavirus NL63 in primary human renal proximal tubular epithelial cells obtained from a commercial supplier, and confirmation of its replication in two different types of human primary kidney cells

doi: 10.1186/1743-422X-10-213

Figure Lengend Snippet: HCoV-NL63/RPTEC/2004 titers in cultured cells. [ A ]. Virus titers seven days post-infection of indicator cells infected with HCoV-NL63/RPTEC/2004 pp A. Titers were obtained from free virus in spent media; plaque assays were performed in CaCo-2 cells. Average virus titers (PFU/ml, mean of 3 measurements) were: LLC-MK2 cells, 3.2 × 10 5 ; Vero E6 cells, 2.3 × 10 4 ; HEK-293 cells, 5.9 × 10 4 ; CV-1 cells, 1.6 × 10 4 ; MDCK-NBL cells, 4.3 × 10 3 ; MDCK-London cells, 4.1 × 10 3 ; Mv1 Lu cells, 6.9 × 10 3 ; WI-38 cells, none detected. [ B] . Virus production over nine days by NL63/RPTEC/2004 pp D in LLC-MK2 cells. Titers (PFU/ml) peaked on day 6, and remained in the low 10 5 range thereafter until day 9 p.i. (last day of measurement).

Article Snippet: Cell lines A549 (CCL-185), BHK-21 (CCL-10), CaCo-2 (HTB-37), CV-1 (CCL-70), HEK-293 (CRL-1573), LLC-MK2 (CCL-7), MDCK, (CCL-34), Mv1 Lu (CCL-64), NIH/3 T3 (CRL-1658), Vero E6 (CRL-1586), and WI-38 (CCL-75) were obtained from the ATCC (Manassas, VA), and along with MDCK-London cells, were propagated as monolayers at 37°C and 5% CO 2 in Dulbecco's Modified Eagle's Medium (DMEM) (Mediatech, Inc., Manassas, VA) or Eagle’s Minimal Essential Medium (EMEM) (Invitrogen Corp., Carlsbad, CA), as appropriate per cell line.

Techniques: Cell Culture, Infection

Journal: PLoS ONE

Article Title: The Histone H2A Variant MacroH2A1 Does Not Localize to the Centrosome

doi: 10.1371/journal.pone.0017262

Figure Lengend Snippet: A . WI-38 cells were transfected with GFP-macroH2A1 and immune-stained for γ-Tubulin as a marker of the centrosome (Red). B . WI-38 cells were co-stained for γ-Tubulin (red) and macroH2A1-NHR antibody (Green). DNA stained with DAPI (Blue). Bar indicates scale of 10 µm.

Article Snippet: HEK293T cells were cultured in DMEM medium and Tert-immortalized WI-38 cells ( , ATCC number CCL-75) in MEM Both media were supplemented with 10% fetal calf serum (FCS), penicillin (50 mg/ml), streptomycin (50 mg/ml) and 2 mM (L-Glutamine).

Techniques: Transfection, Staining, Marker

Journal: PLoS ONE

Article Title: The Histone H2A Variant MacroH2A1 Does Not Localize to the Centrosome

doi: 10.1371/journal.pone.0017262

Figure Lengend Snippet: A . Western blot verifying KD efficiency. B . WI-38 were transduced with either a scrambled vector or a macroH2A1 KD Cells were then subjected to immunofluorescence using antibodies against γ-Tubulin (red) and macroH2A1-NHR (green). DNA stained with DAPI (Blue). Bar indicates scale of 10 µm.

Article Snippet: HEK293T cells were cultured in DMEM medium and Tert-immortalized WI-38 cells ( , ATCC number CCL-75) in MEM Both media were supplemented with 10% fetal calf serum (FCS), penicillin (50 mg/ml), streptomycin (50 mg/ml) and 2 mM (L-Glutamine).

Techniques: Western Blot, Transduction, Plasmid Preparation, Immunofluorescence, Staining

Journal: PLoS ONE

Article Title: The Reverse Transcription Inhibitor Abacavir Shows Anticancer Activity in Prostate Cancer Cell Lines

doi: 10.1371/journal.pone.0014221

Figure Lengend Snippet: (A) Endogenous RT activity was detected in PC3, LNCaP and WI-38 cells as described in ; (+) positive control reaction with commercial RT. (B) Prostate cancer and normal human fibroblast WI-38 cells were treated with ABC at the concentration of 15 and 150 µM and cultured at the indicated time points. Data shown are representative of at least three independent experiments; bars, ± SD.

Article Snippet: The human prostate cancer cell lines PC3 (ATCC CRL-1435) and LNCaP (ATCC CRL-1740) and the non-transformed human fibroblast cell line WI-38 (ATCC CCL-75) were cultured according to the ATCC recommendations.

Techniques: Activity Assay, Positive Control, Concentration Assay, Cell Culture