Mv1lu

Manufactured by American Type Culture Collection

Sourced in United States

The Mv1Lu product is a cell line derived from African green monkey kidney cells. This cell line is commonly used in research and development applications.

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Mv1Lu cells (CCL-64) (2 citations)

7 protocols using mv1lu

Cell culture conditions for coronavirus

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The cell lines used in this study were purchased from ATCC: MRC-5 (human lung epithelial cell; ATCC: CCL-171), HCT-8 (human ileocecal adenocarcinoma cell; ATCC: CCL-244), Mv1Lu (mink lung epithelial cell; ATCC: CCL-64), RD (human rhabdomyosarcoma cell; ATCC: CCL-136), LLC-MK2 (rhesus monkey kidney cell; ATCC: CCL-7), Vero (African green monkey kidney cell; ATCC: CCL-81). HCoV-229E (Catalog No. FR-303) and OC43 (Catalog No. FR-302) were obtained from International Reagent Resources (IRR) followed by propagation in MRC-5 and HCT-8 cells for 3–5 days post infection, respectively.
Medium for MRC-5, Mv1Lu, RD, LLC-MK2, and Vero cells was DMEM (Cat# 112-300; Quality Biological, Gaithersburg, MD, USA) supplemented with 10% FBS, 1× MEM nonessential amino acids (NEAA; Cat# 25-025; Corning, Corning, NY, USA), 1 mM of sodium pyruvate (Cat# 25-000; Corning, Corning, NY, USA), 2 mM of L-glutamine (Cat# 25-005; Corning, Corning, NY, USA), and 1× penicillin and streptomycin (Cat#30-002; Corning, Corning, NY, USA) except HCT-8 which was cultured with RPMI 1640 (Cat# 112-025; Quality Biological, Gaithersburg, MD, USA) containing 10% FBS.

Bracci N., Pan H.C., Lehman C., Kehn-Hall K, & Lin S.C. (2020). Improved plaque assay for human coronaviruses 229E and OC43. PeerJ, 8, e10639.

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Establishment and Characterization of Cell Lines

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293T, MDBK (B. taurus), OHH1.K (O. hemionus), OA3.Ts (O. aries), Ch1 Es (C. hircus), ST (S. scrofa), NZP-29 (O. dammah), MDCK (C. lupus familiaris), CRFK (F. catus), Mv1 Lu (N. vison), FoLu (U. cineroargenteus) and NZP-12 (C. obscurus) cells were obtained from ATCC (Manassas, VA). Ferret (M. putorius furo) MA139 cells were obtained from Dr. Janet Hartley (NIAID, Bethesda, MD). Each cell line was grown in DMEM (Thermo Fisher, Waltham, MA) supplemented with 1% penicillin and 100 μg/mL streptomycin and 10% fetal bovine serum.
The PCR 2.1 TOPO and pcDNA-V5-TOPO plasmids were purchased from Thermo Fisher. The pLNCX2 and pVSV-G plasmids were obtained from Clontech (Mountain View, CA). The pCMVgagpol plasmid which encodes the gagpol gene of the Moloney murine leukemia virus was obtained from Cell Biolabs (San Diego, CA). The pcDNA 3.1 plasmid with a C-terminal HA tag was a kind gift from Dr. Klaus Strebel (NIAID, Bethesda, MD).
Anti-HA and anti-V5 mouse monoclonal antibodies were obtained from Thermo Fisher. Anti-Beta-actin rabbit polyclonal antibody was obtained from Sigma-Aldrich (St. Louis, MO). Goat anti-rabbit-HRP and rabbit anti-mouse-HRP antibodies were purchased from Southern Biotech (Birmingham, AL). G418 sulfate was obtained from Corning (New York, NY).

Simpson J., Kozak C.A, & Boso G. (2022). Cross-species transmission of an ancient endogenous retrovirus and convergent co-option of its envelope gene in two mammalian orders. PLOS Genetics, 18(10), e1010458.

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Optimizing Cellular Visualization Techniques

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The mink lung epithelial cell line, Mv1Lu (ATCC, CCL64, Manassas, VA), was obtained from R. W. Holley and maintained as monolayer cultures in Dulbecco-Vogt's medium (DMEM) containing 0.45 % glucose, 10 % calf serum, 10 units/ml penicillin and 10 µg/ml streptomycin. Chinese hamster ovary (CHO) cells were maintained as monolayer cultures in F12 medium, 10 % calf serum, 10 units/ml penicillin and 10 µg/ml streptomycin. Both cell lines were cultured at 37 °C, in a water-saturated atmosphere with 10 % CO₂ in air.
Yeast cells were cultured at 30 °C in SD-uracil medium containing 2 % glucose for the growth period until they reach OD⁶⁰⁰ 1.0 at which time they were incubated with the aptamer ligands (either with DFHBI or PFP-DFHBI) for 60 minutes prior to imaging. Yeast strains used in this study were: BY4735: MATα ade2Δ::hisG his3Δ200leu2Δ0met15Δ0trp1Δ63ura3Δ0::ppGAL1.MCS16.pYES2/JJR1.2, which expresses the control RNA, BY4735:MATαade2Δ::hisGhis3Δ200leu2Δ0met15Δ0trp1Δ63ura3Δ0[pp5S.5SrRNA.tRNA(Lys)aSpinachtRNA(Lys)Term.pYES2/JJR167.1], which expresses Spinach RNA in a tRNA cassette, BY4735:MATαade2Δ::hisGhis3Δ200leu2Δ0met15Δ0trp1Δ63ura3Δ0pp5S.5SrRNA.tRNA(Lys)a2xSpinach2-tRNA(Lys)Term. pYES2/JJR300.2, which expresses two tandem Spinach2 aptamers and BY4735:MATαade2Δ::hisGhis3Δ200leu2Δ0met15Δ0trp1Δ63ura3Δ0::pGAL1a6xPDC::pJJR1, which expresses the 6xPDC IMAGEtag [9 (link)].

Ilgu M., Ray J., Bendickson L., Wang T., Geraskin I., Kraus G.A, & Nilsen-Hamilton M. (2015). Light-up and FRET aptamer reporters; evaluating their applications for imaging transcription in eukaryotic cells. Methods (San Diego, Calif.), 98, 26-33.

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Generating MyoVa deficient A549 cells

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Mv1Lu, A549, HepG2, Clone 9 and HEK293 cells were obtained from ATCC (Manassas, VA). The PAI-1 promotor stable clone of Mv1Lu cells (MLECs-Clone 32) was a gift from Dr. Jung San Huang from Saint Louis University. The cells were cultured at 37 °C in 5% CO₂ in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Waltham, MA, USA) containing 5% FBS (Life Technologies, Waltham, MA, USA) with 50 μg/mL of streptomycin. To generate a stable MyoVa deficiency cell line, HEK293T cells were used to produce indicated lentiviral particles through the viral packaging process. The supernatant containing the virus was collected and filtered. Subsequently, A549 cells were infected with lentiviral particles. Lentiviral particles containing MyoVa (TRCN0000018214) or control shRNA lentiviral particles (TRCN0000018214) were purchased from the National RNAi Core Facility of Academic Sinica (Taiwan). The infected A549 cells were selected using 2–4 μg/mL of puromycin for 2 weeks.

Shih-Wei W., Chih-Ling C., Kao Y.C., Martin R., Knölker H.J., Shiao M.S, & Chen C.L. (2018). Pentabromopseudilin: a myosin V inhibitor suppresses TGF-β activity by recruiting the type II TGF-β receptor to lysosomal degradation. Journal of Enzyme Inhibition and Medicinal Chemistry, 33(1), 920-935.

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Diverse Cell Lines for Viral Infection Studies

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We obtained MDCK-Atlanta, MDCK-London, and MDCK-SIAT1 cells from International Reagent Resources (https://www.internationalreagentresource.org) and MDCK-hCK cells from the University of Wisconsin–Madison (https://www.wisc.edu). We obtained MDCK-NBL2, Vero E6, CV-1, A549, Crandell-Rees Feline Kidney (CRFK) cells, Mv1Lu, RD, Hep-2c, HeLa, and L20B cells from American Type Culture Collection (https://www.atcc.org); these cells were maintained at Division of Scientific Resources, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention (Atlanta, GA, USA). We obtained chicken embryo fibroblasts from Charles River Laboratories (https://www.criver.com). We obtained an additional 25 cell lines (Table 1) from Quidel Corporation (https://www.quidel.com); these lines were preseeded in 24-well plates, except for CRFK and rhesus monkey kidney cells, which were obtained in T-75 flasks and seeded into 24-well plates in the laboratory 1 day before infection.

Wang L., Fan X., Bonenfant G., Cui D., Hossain J., Jiang N., Larson G., Currier M., Liddell J., Wilson M., Tamin A., Harcourt J., Ciomperlik-Patton J., Pang H., Dybdahl-Sissoko N., Campagnoli R., Shi P.Y., Barnes J., Thornburg N.J., Wentworth D.E, & Zhou B. (2021). Susceptibility to SARS-CoV-2 of Cell Lines and Substrates Commonly Used to Diagnose and Isolate Influenza and Other Viruses. Emerging Infectious Diseases, 27(5), 1380-1392.

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Isolation and Culture of Dental Stem Cells

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hDSCs and mouse pulp cells from DSPP^CreTGF-βRII^fl/fl mice were isolated from the pulp from tooth samples obtained, following Institutional Review Board approval at the Children’s Hospital Boston, as previously described (44 (link)). Briefly, tooth specimens were dissected aseptically and incubated with 4 ml of 0.25% trypsin-EDTA (Life Technologies) at 37°C for 30 min. After neutralization with 4 ml of complete medium, solutions were pipetted vigorously to release cells and then passed through a cell strainer (70 μm, Corning), and resulting cells were cultured in complete medium supplemented with ascorbic acid (100 μM) and β-mercaptoethanol (50 μM) (both from Sigma). Mv1Lu (mink lung epithelial cells) (a gift from D. Rifkin, New York University, Langone Medical Center), MDPC-23 (mouse dental papilla cells) (a gift from T. Bottero, University of Michigan), D1 (mouse mesenchymal stem cells), and 7F2 (mouse osteoblasts) (both from American Type Culture Collection) cells were cultured in complete medium composed of 10% FBS, Dulbecco’s modified Eagle’s medium, GlutaMAX, and penicillin (100 U/ml)–streptomycin (100 μg/ml) (all from Gibco, Life Technologies) in a 37°C incubator with 5% CO₂. MEFs, stably transfected with either full-length or ROS-insensitive (M253A) LTGF-β1, were maintained in growth medium supplemented with G418 (Gibco, Life Technologies).

Arany P.R., Cho A., Hunt T.D., Sidhu G., Shin K., Hahm E., Huang G.X., Weaver J., Chen A.C., Padwa B.L., Hamblin M.R., Barcellos-Hoff M.H., Kulkarni A.B, & Mooney D.J. (2014). Photoactivation of Endogenous Latent Transforming Growth Factor–β1 Directs Dental Stem Cell Differentiation for Regeneration. Science translational medicine, 6(238), 238ra69.

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Mammalian Virus Isolation Using Cell Lines

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A variety of cell lines were tested to improve the chances of isolating mammalian viruses, as some viruses replicate in particular cell lines. Cell lines A549 (CCL-185), BHK-21 (CCL-10), HeLa (CCL-2), LLC-MK2 (CCL-7), MDCK, (CCL-34), Mv1 Lu (CCL-64), Neuro-2a (CCL-131), NIH/3 T3 (CRL-1658), and Vero E6 (CRL-1586) were obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA), and propagated as monolayers at 37°C and 5% CO₂ in Dulbecco's Modified Eagle's Medium (DMEM) (Mediatech, Inc., Manassas, VA, USA) or Eagle's Minimal Essential Medium (EMEM) (Invitrogen, Carlsbad, CA, USA), as appropriate per cell line. DMEM and EMEM were supplemented with 2 mM L-Alanyl-L-Glutamine (GlutaMAX, Invitrogen, Carlsbad, CA, USA.), antibiotics [PSN; 50 µg/ml penicillin, 50 µg/ml streptomycin, 100 µg/ml neomycin (Invitrogen, Carlsbad, CA, USA)], and 10% (v/v) low IgG, heat-inactivated gamma-irradiated fetal bovine serum (HyClone, Logan, UT, USA). EMEM was also supplemented with sodium pyruvate (Invitrogen Corp.) and non-essential amino acids (Hyclone, Logan, UT, USA).

Sayler K.A., Barbet A.F., Chamberlain C., Clapp W.L., Alleman R., Loeb J.C, & Lednicky J.A. (2014). Isolation of Tacaribe Virus, a Caribbean Arenavirus, from Host-Seeking Amblyomma americanum Ticks in Florida. PLoS ONE, 9(12), e115769.

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