Comparison of the continuous cell line 293 with human embryo kidney cells and human embryo fibroblast cells for the cultivation of ocular viruses.

Yirrell, David; Roome, A. P. C. H.; Darville, J. M.; Ashley, C. R.; Harbour, J.

doi:10.1136/jcp.36.9.996

Cited by 7 publications

(3 citation statements)

References 4 publications

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“…Conjunctival swabs were taken for virus isolation from the lower fornix of each eye, placed in antibiotic-containing milk-saline transport medium and posted to the Department of Virology, Bristol Public Health Laboratory, where they usually arrived within 24 h. On receipt they were inoculated into primary or secondary human embryo kidney cell cultures (HEK) and into the continuous human embryo kidney-derived cell line 293 (Graham el al. 1977), which has been shown to be sensitive to a variety of ocular adenoviruses including type 8 (Yirrell et al 1983). Identification of adenoviruses was by solid phase immune electron microscopy using the protein A capture method (SPIEM) (Svensson & von Bonsdorff, 1982).…”

Section: Virus Isolation and Serologymentioning

confidence: 99%

Adenovirus type 8 keratoconjunctivitis – an outbreak and its treatment with topical human fibroblast interferon

Reilly

Dhillon²,

Nkanza³

et al. 1986

J. Hyg.

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An outbreak of keratoconjunctivitis is described which involved at least 186 people; adenovirus type 8 was identified in 50 of the cases. Topical human fibroblast interferon was assessed in a double-blind, placebo-controlled study in which 34 patients participated. Seventeen of the 34 trial patients yielded adenovirus type 8; three were infected with adenovirus type 7. The outbreak was curtailed by control of infection measures: principally careful hand-washing by medical personnel between cases and by discouraging attendance of new cases at the Eye Infirmary. Consequently the trial numbers are small. In addition there was a wide interpatient variation in the severity of infection. Therefore it was not possible to make any statistically valid conclusions concerning the recovery rate of patients receiving interferon or placebo.

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Section: Virus Isolation and Serologymentioning

confidence: 99%

Adenovirus type 8 keratoconjunctivitis – an outbreak and its treatment with topical human fibroblast interferon

Reilly

Dhillon²,

Nkanza³

et al. 1986

J. Hyg.

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“…Morfolojisi epitel olan HEK293 hücre hattı 1973 yılında normal insan embriyonik böbrek hücrelerinin Adenovirüs-5 ile transforme edilmesiyle üretilmiştir (12). Hücrelerin ışık mikroskobundaki morfolojisi birbirlerine ağımsı bağlantılar ile bağlanmış yassı adherent tipindedir.…”

Section: Hücre Kültürüunclassified

The Effect of Endoplasmic Reticulum Stress Activator-Tunicamycin on HEK293 Cells

Kalkan¹,

Vural²,

Kömürcü-Bayrak³

2019

Experimed

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ÖZ ABSTRACT ORİJİNAL ARAŞTIRMA Cite this article as: Kalkan MA, Vural B, Kömürcü Bayrak E. The Effect of Endoplasmic Reticulum Stress Activator-Tunicamycin on HEK293 Cells. Experimed 2018; 8(3): 71-8.Amaç: Endoplazmik Retikulum (ER), protein katlanması ve işlenmesi yanı sıra Ca +2 ve glikojen deposu ve hücre membran lipidlerinin biyogenezi gibi fonksiyonlara sahiptir. Hipoksi, viral enfeksiyonlar, yanlış protein katlanmaları, protein birikimleri ve bazı kimyasallar hücrede, ER stresine sebep olarak katlanmamış protein yanıtı (UPR) sistemlerini devreye sokmaktadır. Bu kimyasallardan biri olan Tunikamisin, asparajin-bağlı glikoproteinlerin sentezinin ilk adımında görev alan GPT (GlcNAc fosfotransferaz) enzimini inhibe ederek hücrede ER stresine sebep olan bir antibiyotiktir. Bu çalışmada, ER stresini in vitro araştırmak için en uygun deney şartlarının belirlenmesi amaçlanmıştır.Gereç ve Yöntem: HEK293 hücre hattı uygun kültür koşullarında çoğaltılarak 5 farklı dozda (0,5-1-2-5 ve 10 µg/mL), 12 saatlik Tunikamisin uygulanmıştır. 1-2-4-8 ve 12 saatlik sürelerle ışık mikroskobunda morfolojik değerlendirilmeleri yapılmıştır. Normal HEK293 hücreleri ile 0,5 ve 1 µg/mL Tunikamisin uygulanan hücrelerden total RNA izolasyonu ve cDNA sentezi yapılmıştır. Tunikamisin uygulamasının ER stres açısından değerlendirilmesi için UPR sisteminden seçilen CHOP, BiP, GADD34, ATF4, EDEM1, ASK1, XBP1 (spliced, unspliced ve total), TRAF2, HERP, PDIA4 genlerinin qRT-PCR yöntemi ile gen ekspresyonları normal hücrelere göre kıyaslanmıştır. Bulgular: Tunikamisin uygulamasından 1-2-4-8 ve 12 saatlik ışık mikroskobunda HEK293 hücrelerinin morfolojilerinin sadece 0,5 ve 1 µg/ mL konsantrasyonlarda korunduğu gözlenmiştir. Diğer dozlarda ise daha ilk saatten hücrelerin morfolojik yapılarının bozulduğu gözlenmiştir. qRT-PCR yöntemi ile UPR-ilişkili genlerden BiP, ASK1, PDIA4, s-x-BP1, us-XBP1, t-XBP1, HERP ve CHOP rölatif kantitasyon değerlerinde Tunikamisin uygulanmış HEK293 hücrelerin normal hücrelere göre anlamlı bir artış saptanmıştır. ASK1 ile aynı yolaktan olmasına rağmen TRAF2 geni rölatif kantitasyon değerinde azalma gözlenmiştir. Sonuç: HEK293 hücrelerinde ER stresinin oluşturulabilmesinde 0.5 ve 1 µg/mL'lik Tunikamisin uygulaması uygun bulunmuştur. Daha yüksek dozların HEK293 hücrelerini apoptoza yönlendirdiği düşünülmektedir.Anahtar Kelimeler: Endoplazmik retikulum stresi, tunikamisin, HEK293 hücreleri, katlanmamış protein yanıtı Objective: Among the functions of the Endoplasmic Reticulum (ER) are many different reactions that include the regulation of protein folding and modifications in the lumen, as well as the use of Ca +2 and glycogen storage, the biogenesis of cell membrane lipids. ER homeostasis becomes unbalanced and is recognized as ER stress by the cell. It triggers Unfolded Protein Responce (UPR) systems. Hypoxia, viral infections, unfolded protein accumulation, and some chemicals cause ER stress. Among the chemicals, tunicamycin is an antibiotic known to induce ER stress in the cell by inhibiting the enzyme GlcNAc phosphotran...

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“…Considering the technical difficulty to establish a convenient cellular stratification system to follow step by step the progress of the infection, most of the studies on the effects of HPV gene expression on epithelial differentiation process have been performed on carcinoma derived cell lines [36–38], or immortalized keratinocytes from several tissues [39–41]. …”

Section: Introductionmentioning

confidence: 99%

HPV16-E2 protein modifies self-renewal and differentiation rate in progenitor cells of human immortalized keratinocytes

Domínguez-Catzín

Reveles-Espinoza

Sánchez-Ramos

et al. 2017

Virol J

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BackgroundCervical cancer is the fourth cause of death worldwide by cancer in women and is a disease associated to persistent infection with human papillomavirus (HPV), particularly from two high-risk types HPV16 and 18. The virus initiates its replicative cycle infecting cells located in the basal layer of the epithelium, where a small population of epithelial stem cells is located performing important functions of renewal and maintenance of the tissue. Viral E2 gene is one of the first expressed after infection and plays relevant roles in the replicative cycle of the virus, modifying fundamental processes in the infected cells. Thus, the aim of the present study was to demonstrate the presence of hierarchic subpopulations in HaCaT cell line and evaluate the effect of HPV16-E2 expression, on their biological processes.MethodsHaCaT-HPV16-E2 cells were generated by transduction of HaCaT cell line with a lentiviral vector. The α6-integrin-CD71 expression profile was established by immunostaining and flow cytometric analysis. After sorting, cell subpopulations were analyzed in biological assays for self-renewal, clonogenicity and expression of stemness factors (RT-qPCR).ResultsWe identified in HaCaT cell line three different subpopulations that correspond to early differentiated cells (α6-integrindim), transitory amplifying cells (α6-integrinbri/CD71bri) and progenitor cells (α6-integrinbri/CD71dim). The last subpopulation showed stem cell characteristics, such as self-renewal ability, clonogenicity and expression of the well-known stem cell factors SOX2, OCT4 and NANOG, suggesting they are stem-like cells. Interestingly, the expression of HPV16-E2 in HaCaT cells changed its α6-integrin-CD71 immunophenotype modifying the relative abundance of the cell subpopulations, reducing significantly the percentage of α6-integrinbri/CD71dim cells. Moreover, the expression of the stem cell markers was also modified, increasing the expression of SOX2 and NANOG, but decreasing notably the expression of OCT4.ConclusionsOur data demonstrated the presence of a small subpopulation with epithelial “progenitor cells” characteristics in the HaCaT cell line, and that HPV16-E2 expression on these cells induces early differentiation.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-017-0736-2) contains supplementary material, which is available to authorized users.

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Comparison of the continuous cell line 293 with human embryo kidney cells and human embryo fibroblast cells for the cultivation of ocular viruses.

Cited by 7 publications

References 4 publications

Adenovirus type 8 keratoconjunctivitis – an outbreak and its treatment with topical human fibroblast interferon

Adenovirus type 8 keratoconjunctivitis – an outbreak and its treatment with topical human fibroblast interferon

The Effect of Endoplasmic Reticulum Stress Activator-Tunicamycin on HEK293 Cells

HPV16-E2 protein modifies self-renewal and differentiation rate in progenitor cells of human immortalized keratinocytes

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