Wi 38 va 13

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WI-38 VA-13 is a cell line derived from human embryonic lung fibroblasts. It is a widely used model system for in vitro studies.

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WI-38 (184 citations) Normal lung WI-38 VA-13 subclone 2RA cells (2 citations)

4 protocols using wi 38 va 13

Characterization of Cell Lines and Viruses

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The 293 T, A549, Huh-7, MDCK, Vero E6, WI-38, WI-38 VA-13, and BEAS-2B cells were obtained from ATCC and maintained in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) supplemented with 10% fetal bovine serum (FBS). The Calu-3 cells (ATCC) were maintained in DMEM supplemented with 20% FBS. NCI-H292 cells (ATCC) were maintained in RPMI-1640 (Gibco) supplemented with 20% FBS. Normal human bronchial epithelial cells (NHBE) cells (ATCC) were maintained in airway epithelial cell basal medium (ATCC PCS300030) supplemented with a Bronchial/Tracheal Epithelial Cell Growth Kit (ATCC PCS300040). All cells were incubated at 37 °C in 5% CO₂.
The A/WSN/33 virus was generated by reverse genetics as previously described.^{29 (link)} H1N1 (A/Sichuan/01/2009) and H3N2 (A/Donghu/312/2006) were kindly provided by the Influenza Center in China CDC. HRV3 and HRSV were purchased from ATCC and stocked accordingly. HPIV was obtained from Professor Mingzhou Chen, Wuhan University. The SARS-CoV-2 live virus (strain IVCAS 6.7512) was provided by the National Virus Resource, Wuhan Institute of Virology, Chinese Academy of Sciences.

Bai L., Zhao Y., Dong J., Liang S., Guo M., Liu X., Wang X., Huang Z., Sun X., Zhang Z., Dong L., Liu Q., Zheng Y., Niu D., Xiang M., Song K., Ye J., Zheng W., Tang Z., Tang M., Zhou Y., Shen C., Dai M., Zhou L., Chen Y., Yan H., Lan K, & Xu K. (2021). Coinfection with influenza A virus enhances SARS-CoV-2 infectivity. Cell Research, 31(4), 395-403.

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Comprehensive Cell Line Culturing Protocol

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All cell lines were cultured in a humidified incubator at 37°C with 5% CO₂. The following cell lines were obtained from ATCC: HEK 293 T, WI-38, WI-38 VA 13, Daudi, EB1, HS604T, HS616T, HuT 102, MC116, Namalwa, H1963, H196, H209, H526, H524, H82, H69, Raji, SW1271, and TO175T. DEL, L428, SU-DHL-10, WSU-DLCL2, Jurkat, DND41 and SR768 were purchased from DSMZ. A4/Fuk was obtained from JCRB Cell Line Bank. OCI-Ly3 was a kind gift from Dr. Mark Minden (University Health Network Toronto). The human lung fibroblast cell lines WI-38 and WI-38 VA13 were routinely cultured in EMEM supplemented with 10% Fetal Bovine Serum (FBS). SCLC cell lines H1963, H196, SW1271, H209, H526, H524, H82 and H69 were maintained in RPMI 1640 with 10% FBS. The lymphoma cell lines EB1, Daudi, Raji, A4/Fukuda, Jurkat, DND41, WSU-DLCL2, DEL, HUT102, Namalwa and L428 were cultured in RPMI 1640 with 10% FBS. MC116 and SU-DHL-10 cells were maintained in RPMI 1640 with 20% FBS. OCI-Ly3 cells were cultured in IMDM with 20% FBS. The SR786 cell line was cultured in RPMI 1640 with 15% FBS. HS604T cells were maintained in DMEM with 10%FBS and 2 mM Glutamine. HS616T and TO175T cell lines were cultured in DMEM with 10% FBS. HEK 293 T cells were maintained in DMEM with 10% FBS. All cell lines were maintained with a cocktail of penicillin and streptomycin (Gibco).

Zhou Z., Patel M., Ng N., Hsieh M.H., Orth A.P., Walker J.R., Batalov S., Harris J.L, & Liu J. (2014). Identification of synthetic lethality of PRKDC in MYC-dependent human cancers by pooled shRNA screening. BMC Cancer, 14, 944.

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Characterization of CFSC Hepatic Stellate Cell Lines

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CFSC cells were established and characterized over thirty years ago in the laboratory of Marcus Rojkind [7 (link),8 (link)]. It is listed in the Cellosaurus database under accession no. CVCL_M104 as a spontaneously immortalized HSC cell line derived from adult male Wistar rats. From this parenteral cell line four clonal cell lines were derived, namely CFSC-2G (CVCL_4U34), CFSC-3H (CVCL_4U35), CFSC-5H (CVCL_4U36), and CFSC-8B (CVCL_4U37). CFSC-2G cells used in this study were routinely propagated in 10 cm Petri dishes and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-Glutamine, 1 mM sodium pyruvate, 1 × Penicillin/Streptomycin, and 1 × non-essential amino acids. Medium exchange was conducted every second day and cells were subcultured using Accutase solution (#A6964, Sigma-Aldrich). WI-38 (VA-13) (#CCL-75.1, ATCC, Manassas, VA, USA) and HSC-T6 cells [10 (link)] were cultured in the same medium as CFSC-2G cells excluding non-essential amino acids.

Nanda I., Schröder S.K., Steinlein C., Haaf T., Buhl E.M., Grimm D.G, & Weiskirchen R. (2022). Rat Hepatic Stellate Cell Line CFSC-2G: Genetic Markers and Short Tandem Repeat Profile Useful for Cell Line Authentication. Cells, 11(18), 2900.

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Cell Line Authentication and Maintenance

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The cell lines, HeLa, Hep3B, MDA-MB-231, ME-180, HT-1080, H1299, MCF7, U2OS, U937, Huh7, SK-HEP-1, WI38, WI38VA-13, IMR-90, BJ, and BJ-5ta (BJ-T), were purchased from ATCC, and SNU cell lines were purchased from Korean Cell Line Bank (KCLB). All the cells were cultured according to the repository's protocol. To maintain authenticity of the cell lines, frozen stocks were prepared at the second passage from the initial stocks, and every 3 months, a new frozen stock was used. p53^−/− MEFs were prepared by crossing heterozygous p53^+/− mice from the Jackson Laboratory. MEFs were maintained in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FBS and 1% antibiotics.

Kim S.H., Park E.R., Cho E., Jung W.H., Jeon J.Y., Joo H.Y., Lee K.H, & Shin H.J. (2016). Mael is essential for cancer cell survival and tumorigenesis through protection of genetic integrity. Oncotarget, 8(3), 5026-5037.

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