Wi 38

Manufactured by American Type Culture Collection

Sourced in United States

WI-38 is a cell line derived from normal human embryonic lung fibroblasts. It is a widely used cell line in biomedical research and has been extensively characterized.

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WI-38 cells (27 citations) WI-38 (CCL-75) (5 citations) WI-38 fibroblasts (4 citations) WI-38 VA-13 (4 citations) WI-38 (ATCC® CCL-75™) (2 citations) WI-38 lung fibroblasts (2 citations) WI-38 human lung fibroblast cell line (2 citations) WI-38 cells (CCL-75) (2 citations) Normal lung WI-38 VA-13 subclone 2RA cells (2 citations)

184 protocols using wi 38

Melanoma Cell Lines for Drug Screening

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Malignant melanoma cell lines, HM8 (derived from brain metastases) and Melmet 5 (derived from lymph node metastases), were established from melanoma patients at Oslo University Hospital, The Norwegian Radium hospital (Oslo, Norway), as described previously [20] (approved by the Norwegian Research Ethics Committee 2011/2183, S-01252, 2.2007.997). The melanoma cells were stably labeled with green fluorescent protein (GFP) -luciferase (LUC) construct, described previously [21] and kindly provided by Dr. Glenn Merlino (NIH, MD).
Human lung fibroblasts WI-38 were obtained from ATCC (Rockville, MD). Melanoma cells were cultured in RPMI 1640 medium, supplemented with 10% fetal calf serum (FCS) and 2 mM L-Alanyl L-Glutamine (all from Sigma-Aldrich, St. Louis, MO). WI-38 fibroblasts were cultured in EMEM medium (ATCC, Manassas, VA), supplemented with 10% FCS. All cell cultures were maintained at 37°C in a humidified atmosphere containing 5% CO2 and were routinely tested for mycoplasma and cell ID.
The BRAF inhibitor vemurafenib, PI3K inhibitor BKM120 and GSK3 inhibitor AR-A014418 were from Selleck Chemicals (Houston, TX). All drugs were dissolved in DMSO.

Seip K., Jørgensen K., Haselager M.V., Albrecht M., Haugen M.H., Egeland E.V., Lucarelli P., Engebraaten O., Sauter T., Mælandsmo G.M, & Prasmickaite L. (2018). Stroma-induced phenotypic plasticity offers phenotype-specific targeting to improve melanoma treatment. Cancer letters, 439.

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Culturing Human Lung Cell Lines

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A549 (human lung adenocarcinoma cell line), H1299 (human lung adenocarcinoma cell line), WI-38 (human lung fibroblast cell line), and MRC-5 (normal human fetal lung fibroblast) cell lines were obtained from American Type Culture Collection (Manassas, VA) and cultured in Dulbecco's Modified Eagle's medium (DMEM; for A549 and H1299) or Basal Medium Eagle (BME; for MRC-5 and WI-38) supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin. All cell cultures were maintained at 37 °C in a humidified atmosphere of 5% CO₂.

Wu H.C., Horng C.T., Lee Y.L., Chen P.N., Lin C.Y., Liao C.Y., Hsieh Y.S, & Chu S.C. (2018). Cinnamomum Cassia Extracts Suppress Human Lung Cancer Cells Invasion by Reducing u-PA/MMP Expression through the FAK to ERK Pathways. International Journal of Medical Sciences, 15(2), 115-123.

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Lung Cell Lines and Fibroblasts Culture

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The lung cell lines (WI-38, A549, NCI-H1573, NCI-H661, NCI-H460, NCI-H1975, NCI-H1563, and NCI-H1299) and human lung fibroblasts (WI-38) cells were purchased from American Type Culture Collection (Manassas, VA). WI38 cells were cultured in Minimum Essential Medium (Invitrogen, Carlsbad, CA), A549 cells were cultured in F12 Kaighn’s Medium (Invitrogen, Carlsbad, CA) and NCI-H1573, NCI-H661, NCI-H460, NCI-H1975, NCI-H1563, and NCI-H1299 cells were cultured in RPMI 1640 (Invitrogen, Carlsbad, CA). All media were supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen, Carlsbad, CA), 100 U/ml penicillin and 100 μg/mL streptomycin (Invitrogen, Carlsbad, CA). The cultures were incubated at 37°C in 5% CO₂/95% humidified air. In all cases, treatment was done in basal medium supplemented with 5% heat-inactivated fetal bovine serum.

Nkembo A.T., Amissah F., Ntantie E., Poku R.A., Salako O.O., Ikpatt O.F, & Lamango N.S. (2019). Polyisoprenylated cysteinyl amide inhibitors deplete K-Ras and induce caspase-dependent apoptosis in lung cancer cells. Current cancer drug targets, 19(10), 838-851.

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Culturing Diverse Human Cell Lines

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A549 (human lung adenocarcinoma cell line), H1299 (human lung adenocarcinoma cell line), WI-38 (human lung fibroblast cell line) and HK-2 (human PTC line derived from a healthy kidney) cell lines were obtained from American Type Culture Collection (Manassas, VA, USA) and cultured in either Dulbecco’s modified Eagle’s medium (DMEM; for A549 and WI38), minimum essential medium (for H1299), or DMEM: Nutrient Mixture F-12 (DMEM/F-12; for HK2) supplemented with 10% fetal bovine serum, 2 mM glutamine, 100 U/mL penicillin, and 100 µg/mL streptomycin. All cell cultures were maintained at 37 °C in a humidified atmosphere of 5% CO₂.

Liu C.M., Shen H.T., Lin Y.A., Yu Y.L., Chen Y.S., Liu C.J, & Hsieh Y.H. (2020). Antiproliferative and Antimetastatic Effects of Praeruptorin C on Human Non–Small Cell Lung Cancer through Inactivating ERK/CTSD Signalling Pathways. Molecules, 25(7), 1625.

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Cell Lines for Neuroblastoma Research

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SK-N-AS, SK-N-FI, SK-N-BE(2)-C, SH-EP, SH-SY5Y, IMR-32, H460, A549, DU145, HCT116, HT29, PC3, SKOV3, CHP-134, Kelly, MDA-MB-231, and HEK-293T cells were purchased from ATCC by the Children’s Cancer Institute Australia Tumour Bank, and maintained in Dulbecco’s modified Eagle’s medium (Invitrogen, Australia) with 10% fetal calf serum. The human lung fibroblast cells MRC-5 and WI-38 (ATCC) were cultured in Alpha-MEM media with 10% FCS. Non-MYCN-amplified human neuroblastoma cell lines SH-EP MYCN-3 and SHEP-tet21N, which are genetically modified to overexpress or repress human MYCN cDNA when exposed to doxycycline, were generously supplied by Professor Jason Shohet (Texas Children’s Cancer Center, Houston). The shMYCN SK-N-BE(2)-C cell line are genetically modified to repress human MYCN expression by MYCN shRNA when exposed to doxycycline, and SK-N-BE(2)-C cell line was used as the parental cell line. The shMYCN SK-N-BE(2)-C cell line was derived by GenScript, Piscataway NJ. All cell lines used were authenticated by CellBank Australia (Westmead, NSW), to be free from mycoplasma, and were cultured at 37 °C with 5% CO₂ in a humidifier incubator.

Cheung B.B., Kleynhans A., Mittra R., Kim P.Y., Holien J.K., Nagy Z., Ciampa O.C., Seneviratne J.A., Mayoh C., Raipuria M., Gadde S., Massudi H., Wong I.P., Tan O., Gong A., Suryano A., Diakiw S.M., Liu B., Arndt G.M., Liu T., Kumar N., Sangfelt O., Zhu S., Norris M.D., Haber M., Carter D.R., Parker M.W, & Marshall G.M. (2021). A novel combination therapy targeting ubiquitin-specific protease 5 in MYCN-driven neuroblastoma. Oncogene, 40(13), 2367-2381.

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Lung and Breast Cancer Cell Lines

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H1048 (R273C) and H1299 (p53 null) lung carcinoma cell lines were
purchased from ATCC and maintained in RPMI (Gibco) plus 10% FBS
(Hyclone). WI38 (WT p53) normal lung fibroblasts were purchased from ATCC and
maintained in Eagle’s MEM (ATCC) plus 10% FBS. ABC1 cells
(P278S) (adenocarcinoma) expressing shGFP or shMDM2 (18 (link)) were maintained in MEM plus 10% FBS and
(1 ug/ml) puromycin (Gibco). H1793, H1975 (both R273H; NSCLC), MDA MB
231 (R280K; breast), U2OS (WT p53; osteosarcoma), and 293T cells (WT p53;
human embryonic kidney) were purchased from ATCC and maintained
in DMEM plus 10% FBS.

Frum R.A., Love I.M., Damle P., Mukhopadhyay N.D., Deb S.P., Deb S, & Grossman S.R. (2016). Constitutive Activation of DNA Damage Checkpoint Signaling Contributes to Mutant p53 Accumulation via Modulation of p53 Ubiquitination. Molecular cancer research : MCR, 14(5), 423-436.

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Cell Culture Protocols for Various Cell Lines

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Human lung fibroblast cell line WI-38 (ATCC CCL-75, bought from LGC standards, France), immortal human cell line HeLa (A kind gift from Prof. Marie-Claire de Pauw-Gillet, ULiège), and human liver cancer cell line HepG2 (a kind gift from Prof. Buc-Calderon, UCLouvain) were grown in Dulbecco's modified eagle medium (DMEM, containing 1 mM pyruvate and 1 g/L glucose, Gibco). Immortalized cell line of human colorectal adenocarcinoma cells Caco-2 (a kind gift from Prof. Ana Beloqui Garcia, LDRI, UCLouvain) were grown in DMEM containing 5% non-essential amino acids and 5% glycine. Murine macrophage cell line J774 was kindly provided by Prof. Françoise Van Bambeke, LDRI, UCLouvain. J774 cells were grown in Roswell Park Memorial Institute medium (RPMI 1640 medium with GlutaMAX, Gibco). All the medium were supplemented with 10% FBS and streptomycin-penicillin (100 UI/mL) (Lonza, Belgium). HeLa, HepG2, WI-38, and J774 cells were kept at 37 °C in a 5% CO₂ incubator while Caco-2 cells were kept in a 10% CO₂ incubator.

Khaliq H.A., Ortiz S., Alhouayek M., Neyts T., Muccioli G.G, & Quetin-Leclercq J. (2022). Effect of a methanolic extract of Salvadora oleoides Decne. on LPS-activated J774 macrophages, its in vitro and in vivo toxicity study and dereplication of its chemical constituents. Toxicology Reports, 9, 1742-1753.

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Placental Chorioamniotic Membrane Mesenchymal Stem Cells

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Sample collection and use for research purposes were approved by the Institutional Review Board of CHA General Hospital, Seoul, Korea. All participants provided written informed consent prior to sample collection. The placentas were collected from female subjects who were free of medical, obstetric and surgical complications and who delivered at term (>37 gestational weeks). The CP-MSCs were harvested as previously described (22 (link)). Briefly, the CP-MSCs were collected from the inner side of the chorioamniotic membrane of the placenta and treated with 0.5% collagenase IV (C7661; Sigma-Aldrich, St. Louis, MO, USA) and cultured in MEM-α modification (#32561; Gibco-BRL, NY, USA) supplemented with 10% fetal bovine serum (FBS, #16000-044), 1% penicillin/streptomycin (#15140-122) (all from Invitrogen, Carlsbad, CA, USA), 25 ng/ml human fibroblast growth factor-4 (hFGF-4; #AF-100-31; Peprotech, Inc., Rocky Hill, NJ, USA) and 1 µg/ml heparin (H3149; Sigma-Aldrich).
In addition, the normal fibroblast cell line, WI-38, was purchased from ATCC (#CCL-75, ATCC; Manassas, VA, USA). The cells were cultured in α-MEM containing 10% FBS, 1% penicillin/streptomycin (P/S; 100 mg/ml, Gibco-BRL), and 2 mM L-glutamine (Gibco-BRL).

CHUNG S., RHO S., KIM G., KIM S.R., BAEK K.H., KANG M, & LEW H. (2016). Human umbilical cord blood mononuclear cells and chorionic plate-derived mesenchymal stem cells promote axon survival in a rat model of optic nerve crush injury. International Journal of Molecular Medicine, 37(5), 1170-1180.

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Evaluating Cancer Cell Line Sensitivity

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All the cell lines used in this study (MCF7, MDA-MB-231, MDA-MB-468, DF2 and WI-38) were purchased from ATCC. Cells were grown in DMEM media supplemented with 10% fetal bovine serum, 100 μg/ml gentamycin, and 2 mM l-glutamine. To grow MCF7 cells the medium was also supplemented with 10 μg/ml insulin (NM Penfild, Denmark). Cells were grown at 37°C in 5% CO₂ atmosphere.
Ecdysterone (95% purity, Frog Tech, Russia) was dissolved in DMSO. Thus, DMSO was used as a control for all experiments with ecdysterone (0 μM Ecdy). Doxorubicin (98% purity, Sigma, United States) and 2-DG (98% purity, Sigma, United States) were dissolved in water.

Shuvalov O., Fedorova O., Tananykina E., Gnennaya Y., Daks A., Petukhov A, & Barlev N.A. (2020). An Arthropod Hormone, Ecdysterone, Inhibits the Growth of Breast Cancer Cells via Different Mechanisms. Frontiers in Pharmacology, 11, 561537.

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Cell Viability Assay with 5-MeCIdn

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Human HCT116, MIA PaCa-2, A549, HeLa, WI-38, and U2OS cell lines were obtained from ATCC. All cell lines were grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 37°C with 5% CO₂. For viability studies, cells were treated with a final concentration of 100 μM 5-MeCIdn in the presence of 0.0003% DMSO or with 0.0003% DMSO alone. Every 4 days cells were provided fresh media containing analog/DMSO or DMSO alone. Every 7 days, cells were counted, and 5.0 × 10⁴ cells were transferred into a new plate with fresh media containing nucleotide analog/DMSO or DMSO. For generation of growth curves, the cells were treated with the indicated drugs for 5 days and trypsinized. Viable cells were identified by trypan blue exclusion and counted on a hemocytometer.

Hernandez-Sanchez W., Huang W., Plucinsky B., Garcia-Vazquez N., Robinson N.J., Schiemann W.P., Berdis A.J., Skordalakes E, & Taylor D.J. (2019). A non-natural nucleotide uses a specific pocket to selectively inhibit telomerase activity. PLoS Biology, 17(4), e3000204.

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