Comparative Transcriptomics Analyses of a Vero Cell Line in Suspension Versus Adherent Culture Conditions

2021 ◽  
Author(s):  
Marie-Angélique Sène ◽  
Yu Xia ◽  
Amine A. Kamen
Keyword(s):  
Cell Viability ◽  
Vero Cell ◽  
Cell Line ◽  
Doubling Time ◽  
Epithelial To Mesenchymal Transition ◽  
Vero Cells ◽  
Suspension Cultures ◽  
Mesenchymal Transition ◽  
Vero Cell Line ◽  
Viral Vaccine

Abstract The Vero cell line is the most used continuous cell line for viral vaccine manufacturing. Its anchorage-dependent use renders scaling-up challenging and operations very labor intensive which affects cost effectiveness. Thus, efforts to adapt Vero cells to suspension cultures have been invested but hurdles such as the long doubling time and low cell viability remain to be addressed. In this study, building on the recently published Vero cell line annotated genome, a functional genomics analysis of the Vero cells adapted to suspension is performed to better understand the genetic and phenotypic switches at play during the adaptation of Vero cells from anchorage-dependent to suspension cultures. Results show a downregulation of the epithelial to mesenchymal transition (EMT) pathway, highlighting the dissociation between the adaptation to suspension process and EMT. Surprisingly, an upregulation of cell adhesion components is observed, notably the CDH18 gene, the cytoskeleton pathway, and the extracellular pathway. Moreover, a downregulation of the glycolytic pathway are balanced by an upregulation of the asparagine metabolism pathway, promoting cell adaptation to nutrient deprivation. A downregulation of the adherens junctions and the folate pathways alongside with the FYN gene are possible explanations behind the currently observed low cell viability and long doubling time.

scholarly journals In Vitro Assessment of Gentamicin Cytotoxicity on the Selected Mammalian Cell Line (Vero cells)

2017 ◽  
Vol 1 (1) ◽  
pp. 111-116 ◽  
Author(s):  
Anton Kovacik ◽  
Eva Tvrda ◽  
Diana Fulopova ◽  
Peter Cupka ◽  
Eva Kovacikova ◽  
...  
Keyword(s):  
Cell Viability ◽  
Vero Cell ◽  
Cell Line ◽  
Mammalian Cell ◽  
Vero Cells ◽  
Mammalian Cell Line ◽  
Total Proteins ◽  
African Green Monkey Kidney ◽  
Vero Cell Line

Abstract The aim of this study was to evaluate the in vitro cytotoxicity of different concentrations (500-7500 μg/mL) of gentamicin - GENT (aminoglycoside antibiotic) on the selected mammalian cell line (Vero - cell line from African green monkey kidney). Analysis of the cell morphological changes was microscopically evaluated (magnification × 400). Quantification of Ca, Mg and total proteins was performed using spectrophotometry on device Rx Monza (Randox). Quantification of Na, K and Cl was performed on the automatic analyzer EasyLyte. The cell viability was assessed using the metabolic mitochondrial MTT test. Vero cells were able to survive at concentrations of 500 (89.21 %), 1000 (79.54 %) and 2000 μg/mL (34.59 %). We observed statistically significant decrease of vital cell content at concentrations of 2000, 4500, 7500 μg/mL against control group. Vero cell line slightly reacted to the presence of GENT but total proteins and mineral parameters were not significantly affected. Vero cells were highly sensitive to GENT with a significant decrease of viability at concentrations of 2000 and 4500 μg/mL (P < 0.001). Our data reveal that GENT has a significant cytotoxic and adverse effect on the cell viability.

scholarly journals Cytotoxic and Genotoxic Effects of Fluconazole on African Green Monkey Kidney (Vero) Cell Line

2018 ◽  
Vol 2018 ◽  
pp. 1-7
Author(s):  
Regianne Maciel dos Santos Correa ◽  
Tatiane Cristina Mota ◽  
Adriana Costa Guimarães ◽  
Laís Teixeira Bonfim ◽  
Rommel Rodriguez Burbano ◽  
...  
Keyword(s):  
Comet Assay ◽  
Vero Cell ◽  
Cell Line ◽  
Vero Cells ◽  
Negative Control ◽  
African Green Monkey ◽  
Monkey Kidney ◽  
African Green Monkey Kidney ◽  
Vero Cell Line ◽  
Green Monkey

Fluconazole is a broad-spectrum triazole antifungal that is well-established as the first-line treatment for Candida albicans infections. Despite its extensive use, reports on its genotoxic/mutagenic effects are controversial; therefore, further studies are needed to better clarify such effects. African green monkey kidney (Vero) cells were exposed in vitro to different concentrations of fluconazole and were then evaluated for different parameters, such as cytotoxicity (MTT/cell death by fluorescent dyes), genotoxicity/mutagenicity (comet assay/micronucleus test), and induction of oxidative stress (DCFH-DA assay). Fluconazole was used at concentrations of 81.6, 163.2, 326.5, 653, 1306, and 2612.1μM for the MTT assay and 81.6, 326.5, and 1306μM for the remaining assays. MTT results showed that cell viability reduced upon exposure to fluconazole concentration of 1306μM (85.93%), being statistically significant (P<0.05) at fluconazole concentration of 2612.1μM (35.25%), as compared with the control (100%). Fluconazole also induced necrosis (P<0.05) in Vero cell line when cells were exposed to all concentrations (81.6, 326.5, and 1306μM) for both tested harvest times (24 and 48 h) as compared with the negative control. Regarding genotoxicity/mutagenicity, results showed fluconazole to increase significantly (P<0.05) DNA damage index, as assessed by comet assay, at 1306μM versus the negative control (DI=1.17 vs DI=0.28, respectively). Micronucleus frequency also increased until reaching statistical significance (P<0.05) at 1306μM fluconazole (with 42MN/1000 binucleated cells) as compared to the negative control (13MN/1000 binucleated cells). Finally, significant formation of reactive oxygen species (P<0.05) was observed at 1306μM fluconazole vs the negative control (OD=40.9 vs OD=32.3, respectively). Our experiments showed that fluconazole is cytotoxic and genotoxic in the assessed conditions. It is likely that such effects may be due to the oxidative properties of fluconazole and/or the presence of FMO (flavin-containing monooxygenase) in Vero cells.

scholarly journals Bioreactor production of rVSV-vectored vaccines in Vero cell suspension cultures

2021 ◽  
Author(s):  
Sascha Kiesslich ◽  
Gyoung Kim ◽  
Chun Fang Shen ◽  
Chil-Yong Kang ◽  
Amine Kamen
Keyword(s):  
Vero Cell ◽  
Cell Line ◽  
Virus Disease ◽  
Fixed Bed ◽  
Suspension Cultures ◽  
Suspension System ◽  
Small Scale ◽  
Adherent Cell ◽  
Viral Particles ◽  
Viral Vaccine

The Vero cell line is the most used continuous cell line in viral vaccine manufacturing. This adherent cell culture platform requires the use of surfaces to support cell growth, typically roller bottles or microcarriers. We have recently compared the production of rVSV-ZEBOV on Vero cells between microcarrier and fixed-bed bioreactors. However, suspension cultures are considered superior with regards to process scalability. Therefore, we further explore the Vero suspension system for rVSV-vectored vaccine production. Previously, this suspension cell line was only able to be cultivated in a proprietary medium. Here, we expand the adaptation and bioreactor cultivation to a serum-free commercial medium. Following small scale optimization and screening studies, we demonstrate bioreactor productions of highly relevant vaccines and vaccine candidates against Ebola virus disease, HIV and COVID-19 in the Vero suspension system. rVSV-ZEBOV, rVSV-HIV and rVSVInd-msp-SF-Gtc can replicate to high titers in the bioreactor, reaching 3.87 × 107 TCID50/mL, 2.12 × 107 TCID50/mL and 3.59 × 109 TCID50/mL, respectively. Further, we compare cell specific productivities, and the quality of the produced viruses by determining the ratio of total viral particles to infectious viral particles

scholarly journals Haplotype-resolved de novo assembly of the Vero cell line genome

npj Vaccines ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Marie-Angélique Sène ◽  
Sascha Kiesslich ◽  
Haig Djambazian ◽  
Jiannis Ragoussis ◽  
Yu Xia ◽  
...  
Keyword(s):  
Vero Cell ◽  
Cell Line ◽  
De Novo ◽  
Variant Calling ◽  
High Yield ◽  
Vaccine Production ◽  
Emerging Pathogens ◽  
Structural Variations ◽  
Vero Cell Line ◽  
Viral Vaccine

AbstractThe Vero cell line is the most used continuous cell line for viral vaccine manufacturing with more than 40 years of accumulated experience in the vaccine industry. Additionally, the Vero cell line has shown a high affinity for infection by MERS-CoV, SARS-CoV, and recently SARS-CoV-2, emerging as an important discovery and screening tool to support the global research and development efforts in this COVID-19 pandemic. However, the lack of a reference genome for the Vero cell line has limited our understanding of host–virus interactions underlying such affinity of the Vero cell towards key emerging pathogens, and more importantly our ability to redesign high-yield vaccine production processes using Vero genome editing. In this paper, we present an annotated highly contiguous 2.9 Gb assembly of the Vero cell genome. In addition, several viral genome insertions, including Adeno-associated virus serotypes 3, 4, 7, and 8, have been identified, giving valuable insights into quality control considerations for cell-based vaccine production systems. Variant calling revealed that, in addition to interferon, chemokines, and caspases-related genes lost their functions. Surprisingly, the ACE2 gene, which was previously identified as the host cell entry receptor for SARS-CoV and SARS-CoV-2, also lost function in the Vero genome due to structural variations.

scholarly journals Anti-Cancer Effects of Glaucarubinone in the Hepatocellular Carcinoma Cell Line Huh7 via Regulation of the Epithelial-To-Mesenchymal Transition-Associated Transcription Factor Twist1

2021 ◽  
Vol 22 (4) ◽  
pp. 1700
Author(s):  
Jihye Seo ◽  
Jain Ha ◽  
Eunjeong Kang ◽  
Haelim Yoon ◽  
Sewoong Lee ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Wound Healing ◽  
Transcription Factor ◽  
Cell Migration ◽  
Cell Line ◽  
Intracellular Signaling ◽  
Epithelial To Mesenchymal Transition ◽  
Mesenchymal Transition ◽  
Huh7 Cells ◽  
Anti Cancer

Hepatocellular carcinoma (HCC), the most common type of liver cancer, is a leading cause of cancer-related deaths. As HCC has a high mortality rate and its incidence is increasing worldwide, understanding and treating HCC are crucial for resolving major public health concerns. In the present study, wound healing screening assays were performed using natural product libraries to identify natural chemicals that can inhibit cancer cell migration. Glaucarubinone (GCB) showed a high potential for inhibiting cell migration. The anti-cancer effects of GCB were evaluated using the HCC cell line, Huh7. GCB showed anti-cancer effects, as verified by wound healing, cell migration, invasion, colony formation, and three-dimensional spheroid invasion assays. In addition, cells treated with GCB showed suppressed matrix metalloproteinase activities. Immunoblotting analyses of intracellular signaling pathways revealed that GCB regulated the levels of Twist1, a crucial transcription factor associated with epithelial-to-mesenchymal transition, and mitogen-activated protein kinase. The invasive ability of cancer cells was found to be decreased by the regulation of Twist1 protein levels. Furthermore, GCB downregulated phosphorylation of extracellular signal-regulated kinase. These results indicate that GCB exhibits anti-metastatic properties in Huh7 cells, suggesting that it could be used to treat HCC.

Whole Genome Sequencing and Phylogenetic Analysis of the Rabies Virus Strain Moscow 3253 Adapted to a Vero Cell Line

2020 ◽  
Vol 35 (4) ◽  
pp. 237-242
Author(s):  
Ya. M. Krasnov ◽  
Zh. V. Alkhova ◽  
S. V. Generalov ◽  
I. V. Tuchkov ◽  
E. A. Naryshkina ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Vero Cell ◽  
Whole Genome Sequencing ◽  
Cell Line ◽  
Rabies Virus ◽  
Genome Sequencing ◽  
Virus Strain ◽  
Whole Genome ◽  
Vero Cell Line ◽  
Rabies Virus Strain

scholarly journals Antibacterial Activity, Cytotoxicity, and the Mechanism of Action of Bacteriocin from Bacillus subtilis GAS101

2018 ◽  
Vol 27 (2) ◽  
pp. 186-192 ◽  
Author(s):  
Garima Sharma ◽  
Shweta Dang ◽  
Sanjay Gupta ◽  
Reema Gabrani
Keyword(s):  
Molecular Weight ◽  
Antibacterial Activity ◽  
Bacillus Subtilis ◽  
Vero Cell ◽  
Cell Line ◽  
Mode Of Action ◽  
Proteolytic Enzymes ◽  
Antibiofilm Activity ◽  
Vero Cell Line ◽  
Soil Isolate

Objective: The aim of this study was to purify and characterize bacteriocin from the soil isolate Bacillus subtilis GAS101, and to determine its antimicrobial as well as antibiofilm potential. The purified bacteriocin was further analyzed and evaluated for mammalian cell cytotoxicity and the possible mode of action. Material and Methods: Bacteriocin from B. subtilis GAS101 (an animal husbandry soil isolate) was partially purified and checked for antimicrobial and antibiofilm activity against gram-positive and gram-negative bacteria. The molecular weight of bacteriocin was determined using tricine SDS-PAGE gel. The stability of bacteriocin was investigated at various temperatures and pH levels, and its sensitivity towards 8 enzymes and 6 chemicals was determined. Cytotoxicity analysis was performed on a Vero cell line by a tetrazolium dye-based assay. Scanning electron microscopy (SEM) of bacteriocin-treated bacteria was carried out to determine the possible mode of action. Results: Bacteriocin from B. subtilis GAS101 was a potential inhibitor of both the indicator organisms (Staphylococcus epidermidis and Escherichia coli), and had a molecular weight of approximately 6.5 kDa. An in situ gel assay showed a zone of inhibition corresponding to the estimated protein band size. Bacteriocin was stable and showed antibacterial activity in broad ranges of temperature (30–121°C) and pH (2–12). It was sensitive to 4 proteolytic enzymes, which indicated its proteinaceous nature. Bacteriocin showed > 70% cell viability on the mammalian Vero cell line. SEM depicted that the bacteriocin was able to disrupt the bacterial cell membrane as its probable mode of action. Conclusion: Thermostable and pH-tolerant bacteriocin from B. subtilis GAS101, of about 6.5 kDa, showed broad-spectrum antimicrobial and antibiofilm activity.

scholarly journals Flavonoids from Milletia leucantha and Their Cytotoxicity

2018 ◽  
Vol 13 (8) ◽  
pp. 1934578X1801300
Author(s):  
Uraiwan Sriphana ◽  
Chavi Yenjai ◽  
Siriporn Tungnoi ◽  
Jongjai Srirapa ◽  
Auemporn Junsongduang
Keyword(s):  
Cytotoxic Activity ◽  
Vero Cell ◽  
Cell Line ◽  
Vero Cells ◽  
2D Nmr ◽  
Nmr Techniques ◽  
Vero Cell Lines ◽  
First Time ◽  
Flavonol Derivatives ◽  
Mcf 7

Two known chalcones (1–2), five known flavonol derivatives (3 and 5–8) and one flavone (4) were isolated for the first time from fruits of Millettia leucantha Kurz. In addition, 5 was found for the first time from this genus. Their structures were elucidated on the basis of IR, MS, 1D and 2D NMR techniques. Chalcone 1 exhibited moderate cytotoxicity against the MCF-7 cell line with an IC50 value of 50.93 μM, whereas this compound showed inactive cytotoxicity against Vero cells. Compound 2 exhibited cytotoxicity against the KB, NCI-H187 and Vero cell lines with IC50 values of 63.64, 114.44 and 28.44 μM, respectively. Compound 8 only showed cytotoxic activity against the KB cell line with an IC50 value of 110.23 μM.

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