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Ohio hela cells

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The Ohio HeLa cells are a widely used immortalized cell line derived from human cervical cancer cells. They are a valuable research tool that provides a consistent and renewable source of human cells for various scientific studies and applications.

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Wt balb c, by Jackson ImmunoResearch (1 mentions) Hela cell, by American Type Culture Collection (1 mentions) Amicon ultra 15, by Merck Group (1 mentions) Polyethylene glycol 6000, by Merck Group (1 mentions) Beas 2b cells, by European Collection of Cell Cultures (1 mentions) 1640 medium, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)

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HeLa cells (11 citations)

8 protocols using ohio hela cells

1

Rhinovirus RV16 Infection in HBECs

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The major group rhinovirus RV16 was grown in Ohio HeLa cells (European Collection of Cell Cultures) as previously described [36 (link)] and was obtained from clarified cell lysates. HBECs were infected with RV16 at 1 multiplicity of infection (MOI) [28 (link), 37 (link)] for 1h at room temperature while shaking. Then the virus was removed and fresh BEGM medium containing azithromycin was added. Cell lysates and supernatants were obtained 24h and 48h post infection, respectively, and utilized for gene and protein expression analysis.
Menzel M., Akbarshahi H., Tufvesson E., Persson C., Bjermer L, & Uller L. (2017). Azithromycin augments rhinovirus-induced IFNβ via cytosolic MDA5 in experimental models of asthma exacerbation. Oncotarget, 8(19), 31601-31611.
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2

Rhinovirus Infection of Airway Cells

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The minor group rhinovirus RV1B was grown in Ohio HeLa cells (European Collection of Cell Cultures) as previously described [29 (link)] and was obtained from clarified cell lysates (1.58×107 tissue culture infectious dose (TCID50)/mL). Based on our previous studies we chose an optimal concentration of RV for the infection experiments [10 (link), 12 (link)]. BSMCs were infected with RV1B at 1 multiplicity of infection (MOI) for 1 h at room temperature while shaking. Thereafter, the inoculum was removed and replaced with DMEM containing 1% FBS. In further experiments, BSMCs were stimulated with different PRR agonists such as TLR3 agonist poly(I:C) (10 μg·mL−1), TLR7 agonist imiquimod (10 μg·mL−1) and RIG-I/MDA5 agonist poly(I:C)/LyoVec (0.5 μg·mL−1). Cell lysates and supernatants were collected 24 h post-infection, respectively, and utilised for gene and protein expression analysis, respectively.
Ramu S., Calvén J., Michaeloudes C., Menzel M., Akbarshahi H., Chung K.F, & Uller L. (2020). TLR3/TAK1 signalling regulates rhinovirus-induced interleukin-33 in bronchial smooth muscle cells. ERJ Open Research, 6(4), 00147-2020.
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3

Rhinovirus Infection in BALB/c Mice

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WT BALB/c mice were purchased from Harlan Laboratories and maintained in specific pathogen-free conditions at Imperial College London (UK). Mice aged 6-8 weeks were used for each experiment. All animal experiments were reviewed and approved by the Animal Welfare and Ethical Review Board (AWERB) within Imperial College London and approved by the UK Home Office in accordance with the Animals Act 1986 and the ARRIVE guidelines. Minor-group RV serotype-1b (RV1b) were grown in Ohio HeLa cells (European Collection of Cell Cultures). Viruses were obtained from the American Type Culture Collection and were passaged five times in HeLa cells prior purification6 (link). Viruses were titrated on HeLa cells by standard methods and were inactivated by UV-light exposure at 1.200mJ/cm2 for 30 minutes. WT BALB/c and eosinophil-deficient ΔdblGATA (C.Cg-Gata1tm6Sho/J) BALB/c mice were purchased from The Jackson Laboratory, and mice were housed and bred in specific pathogen-free facilities at the University of Liège (Belgium). Mice aged 6-8 weeks were used and experiments were approved by the IACUC of the University of Liège (approval number 1593).
Toussaint M., Jackson D.J., Swieboda D., Guedán A., Tsourouktsoglou T.D., Ching Y.M., Radermecker C., Makrinioti H., Aniscenko J., Edwards M.R., Solari R., Farnir F., Papayannopoulos V., Bureau F., Marichal T, & Johnston S.L. (2017). Host DNA released by NETosis promotes rhinovirus-induced type 2 allergic asthma exacerbation. Nature medicine, 23(6), 681-691.
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4

Inactivating Rhinovirus for Mouse Studies

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BEAS-2B cells (European Collection of cell cultures) were cultured in RPMI 1640 medium with 10% foetal calf serum (FCS). Rhinovirus serotype A1 was propagated in Ohio HeLa cells (European Collection of cell cultures) by standard methods and inactivated by exposure to UV light at 1200 mJ/cm2 for 30 min. For in vivo use in mouse models, virus was purified using precipitation in 7% polyethylene glycol 6000 (Sigma-Aldrich), 0.5 M NaCl, followed by filtration using a 0.2 μm syringe filter and buffer exchange and concentration in an Amicon Ultra-15 centrifugal filter unit (100 KDa nominal molecular weight limit) (Millipore, USA)33 (link),59 (link).
Singanayagam A., Glanville N., Girkin J.L., Ching Y.M., Marcellini A., Porter J.D., Toussaint M., Walton R.P., Finney L.J., Aniscenko J., Zhu J., Trujillo-Torralbo M.B., Calderazzo M.A., Grainge C., Loo S.L., Veerati P.C., Pathinayake P.S., Nichol K.S., Reid A.T., James P.L., Solari R., Wark P.A., Knight D.A., Moffatt M.F., Cookson W.O., Edwards M.R., Mallia P., Bartlett N.W, & Johnston S.L. (2018). Corticosteroid suppression of antiviral immunity increases bacterial loads and mucus production in COPD exacerbations. Nature Communications, 9, 2229.
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5

PBMCs Rhinovirus-16 Infection Protocol

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PBMCs were cultured in 24-well plates at a cell density of 1×106/mL in RPMI (Roswell Park Memorial Institute)-1640 medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich Co.) and infected with a major group rhinovirus, rhinovirus-16 (multiplicity of infection, 1). Rhinovirus-16 was grown in Ohio Hela cells (European Collection of Cell Cultures) and stocks were generated and titrated [10 (link)]. As a negative control, cells were treated with medium alone. Cells and supernatants were harvested at 12 hours after rhinovirus stimulation.
Kim S.H., Lim K.H., Park H.K., Lee S.Y., Kim S.H., Kang H.R., Park H.W., Chang Y.S, & Cho S.H. (2015). Reduced IRF7 response to rhinovirus unrelated with DNA methylation in peripheral mononuclear cells of adult asthmatics. Asia Pacific Allergy, 5(2), 114-122.
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6

Rhinovirus RV16 infection of HBECs

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Rhinovirus RV16 was amplified in Ohio HeLa cells (European Collection of Cell Cultures) as described previously (24 (link)) and obtained from clarified cell lysates. HBECs were infected with RV16 at 1MOI (TCID50 1.58 × 105) for 1 h at room temperature while shaking. Then virus was removed and fresh culture medium was added. After 48 h cell lysates were collected for subsequent protein expression analysis. A characterization of patients can be found in Table 3.
Menzel M., Ramu S., Calvén J., Olejnicka B., Sverrild A., Porsbjerg C., Tufvesson E., Bjermer L., Akbarshahi H, & Uller L. (2019). Oxidative Stress Attenuates TLR3 Responsiveness and Impairs Anti-viral Mechanisms in Bronchial Epithelial Cells From COPD and Asthma Patients. Frontiers in Immunology, 10, 2765.
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7

Rhinovirus Infection in Human Bronchial Epithelial Cells

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For all experiments cells were used at passage 2–4. HBECs were infected with the major group rhinovirus RV16, grown in Ohio HeLa cells (European Collection of Cell Cultures) as described previously37 (link) and RV16 was obtained from clarified cell lysates. HBECs were infected with RV16 at 1 multiplicity of infection (MOI; TCID50 1.58 × 105) for 1 h at room temperature while shaking. Then the virus was removed and cells were washed with phosphate buffered saline (PBS). Fresh BEGM medium containing azithromycin was added. Eight hours and 24 h post infection cells were lysed and harvested for gene expression analysis and 48 h post infection cells were lysed and harvested for protein expression analysis.
Menzel M., Akbarshahi H., Bjermer L, & Uller L. (2016). Azithromycin induces anti-viral effects in cultured bronchial epithelial cells from COPD patients. Scientific Reports, 6, 28698.
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8

RV-A1 Propagation and Titration

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For the mouse and human cell line studies, minor group RV serotype-A1 (RV-A1) was grown in Ohio HeLa cells (European Collection of Cell Cultures). Infected cells were harvested after 24 hours, and the virus was concentrated and purified and a TCID 50 (50% tissue culture infective dose) assay was performed to assess the viral titer as described previously. 12 12. Bartlett, N.W. • Walton, R.P. • Edwards, M.R. ... For the human primary bronchial epithelial cell (BEC) studies, RV-A1 was propagated in RD-ICAM-1 cells from an in-house stock isolated from clinical samples and sequenced to confirm identity. RV-A1 was titrated by infecting RD-ICAM-1 cells with serially diluted RV-A1, followed by observations of cytopathic effects and TCID 50 assay to assess the viral titer.
Antunes K.H., Singanayagam A., Williams L., Faiez T.S., Farias A., Jackson M.M., Faizi F.K., Aniscenko J., Kebadze T., Chander Veerati P., Wood L., Bartlett N.W., Duarte de Souza A.P, & Johnston S.L. (2023). Airway-delivered short-chain fatty acid acetate boosts antiviral immunity during rhinovirus infection. The Journal of allergy and clinical immunology, 151(2).
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