Complete Genome Sequences, before and after Mammalian Cell Culture, of Zika Virus Isolated from the Serum of a Symptomatic Male Patient from Oaxaca, Mexico

ABSTRACT Zika virus (ZIKV) is an emerging arthropod-borne flavivirus associated with severe congenital malformations and neurological complications. Although the ZIKV genome is well characterized, there is limited information regarding changes after cell isolation and culture adaptation. We isolated, and passaged in Vero cells, ZIKV from the serum of a symptomatic male patient and compared the viral genomes before and after culture. Single nucleotide polymorphisms were characteristic among serum-circulating genomes, while such diversity decreased after cell culture.

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Faculty Opinions recommendation of Zika virus outbreak: a review of neurological complications, diagnosis, and treatment options.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.732666093.793581255 ◽

2020 ◽

Author(s):

Tian Wang

Keyword(s):

Zika Virus ◽

Treatment Options ◽

Neurological Complications ◽

Diagnosis And Treatment

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The P-MAPA immunomodulator partially prevents apoptosis induced by Zika virus infection in THP-1 cells

Current Pharmaceutical Biotechnology ◽

10.2174/1389201021666200602140005 ◽

2020 ◽

Vol 21 ◽

Cited By ~ 1

Author(s):

Morganna C. Lima ◽

Elisa A. N. Azevedo ◽

Clarice N. L. de Morais ◽

Larissa I. O. de Sousa ◽

Bruno M. Carvalho ◽

...

Keyword(s):

Cell Proliferation ◽

Cell Death ◽

Virus Infection ◽

Virus Replication ◽

Zika Virus ◽

Mammalian Cells ◽

Caspase 3 ◽

Neurological Complications ◽

Zika Virus Infection

Background: Zika virus is an emerging arbovirus of global importance. ZIKV infection is associated with a range of neurological complications such as the Congenital Zika Syndrome and Guillain Barré Syndrome. Despite the magnitude of recent outbreaks, there is no specific therapy to prevent or to alleviate disease pathology. Objective: To investigate the role of P-MAPA immunomodulator in Zika-infected THP-1 cells. Methods: THP-1 cells were subjected at Zika virus infection (Multiplicity of Infection = 0.5) followed by treatment with P-MAPA for until 96 hours post-infection. After that, the cell death was analyzed by annexin+/ PI+ and caspase 3/ 7+ staining by flow cytometry. In addition, the virus replication and cell proliferation were accessed by RT-qPCR and Ki67 staining, respectively. Results: We demonstrate that P-MAPA in vitro treatment significantly reduces Zika virus-induced cell death and caspase-3/7 activation on THP-1 infected cells, albeit it has no role in virus replication and cell proliferation. Conclusions: Our study reveals that P-MAPA seems to be a satisfactory alternative to inhibits the effects of Zika virus infection in mammalian cells.

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Protein Purification Techniques

10.1093/oso/9780199636747.001.0001 ◽

2001 ◽

Keyword(s):

Cell Culture ◽

Protein Purification ◽

Mammalian Cell Culture ◽

Scale Up ◽

Analytical Techniques ◽

Structure And Function ◽

Unit Operations ◽

Practical Guidelines ◽

P Proteins ◽

And Function

Proteins are an integral part of molecular and cellular structure and function and are probably the most purified type of biological molecule. In order to elucidate the structure and function of any protein it is first necessary to purify it. Protein purification techniques have evolved over the past ten years with improvements in equipment control, automation, and separation materials, and the introduction of new techniques such as affinity membranes and expanded beds. These developments have reduced the workload involved in protein purification, but there is still a need to consider how unit operations linked together to form a purification strategy, which can be scaled up if necessary. The two Practical Approach books on protein purification have therefore been thoroughly updated and rewritten where necessary. The core of both books is the provision of detailed practical guidelines aimed particularly at laboratory scale purification. Information on scale-up considerations is given where appropriate. The books are not comprehensive but do cover the major laboratory techniques and common sources of protein. Protein Purification Techniques focuses on unit operations and analytical techniques. It starts with an overview of purification strategy and then covers initial extraction and clarification techniques. The rest of the book concentrates on different purification methods with the emphasis being on chromatography. The final chapter considers general scale-up considerations. Protein Purification Applications describes purification strategies from common sources: mammalian cell culture, microbial cell culture, milk, animal tissue, and plant tissue. It also includes chapters on purification of inclusion bodies, fusion proteins, and purification for crystallography. A purification strategy that can produce a highly pure single protein from a crude mixture of proteins, carbohydrates, lipids, and cell debris to is a work of art to be admired. These books (available individually or as a set)are designed to give the laboratory worker the information needed to undertake the challenge of designing such a strategy.

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Miniature Bioreactors for Rapid Bioprocess Development of Mammalian Cell Culture

Jurnal Teknologi ◽

10.11113/jt.v59.1569 ◽

2012 ◽

Vol 59 (1) ◽

Author(s):

Mohd Helmi Sani ◽

Frank Baganz

Keyword(s):

Cell Culture ◽

Mammalian Cell ◽

Process Development ◽

Mammalian Cell Culture ◽

Small Scale ◽

Cell Culture Process ◽

Deep Wells ◽

Working Volume ◽

Culture Process ◽

And Control

At present, there are a number of commercial small scale shaken systems available on the market with instrumented controllable microbioreactors such as Micro–24 Microreactor System (Pall Corporation, Port Washington, NY) and M2P Biolector, (M2P Labs GmbH, Aachen, Germany). The Micro–24 system is basically an orbital shaken 24–well plate that operates at working volume 3 – 7 mL with 24 independent reactors (deep wells, shaken and sparged) running simultaneously. Each reactor is designed as single use reactor that has the ability to continuously monitor and control the pH, DO and temperature. The reactor aeration is supplied by sparging air from gas feeds that can be controlled individually. Furthermore, pH can be controlled by gas sparging using either dilute ammonia or carbon dioxide directly into the culture medium through a membrane at the bottom of each reactor. Chen et al., (2009) evaluated the Micro–24 system for the mammalian cell culture process development and found the Micro–24 system is suitable as scaledown tool for cell culture application. The result showed that intra-well reproducibility, cell growth, metabolites profiles and protein titres were scalable with 2 L bioreactors.

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Identification of contaminating mollicutes in mammalian cell culture as spiroplasma species

Biologicals ◽

10.1016/j.biologicals.2021.04.001 ◽

2021 ◽

Author(s):

Helena Windsor ◽

Christie English

Keyword(s):

Cell Culture ◽

Mammalian Cell ◽

Mammalian Cell Culture

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Production of Newcastle Disease Virus by Vero Cells Grown on Cytodex 1 Microcarriers in a 2-Litre Stirred Tank Bioreactor

Journal of Biomedicine and Biotechnology ◽

10.1155/2010/586363 ◽

2010 ◽

Vol 2010 ◽

pp. 1-7 ◽

Cited By ~ 15

Author(s):

Mohd Azmir Arifin ◽

Maizirwan Mel ◽

Mohamed Ismail Abdul Karim ◽

Aini Ideris

Keyword(s):

Cell Culture ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

High Speed ◽

Virus Titer ◽

Vero Cells ◽

Stirred Tank ◽

Stirred Tank Bioreactor ◽

Maximum Cell

The aim of this study is to prepare a model for the production of Newcastle disease virus (NDV) lentogenic F strain using cell culture in bioreactor for live attenuated vaccine preparation. In this study, firstly we investigated the growth of Vero cells in several culture media. The maximum cell number was yielded by culture of Vero cells in Dulbecco's Modified Eagle Medium (DMEM) which was1.93×106 cells/ml. Secondly Vero cells were grown in two-litre stirred tank bioreactor by using several commercial microcarriers. We achieved the maximum cell concentration about7.95×105 cells/ml when using Cytodex 1. Later we produced Newcastle Disease virus in stirred tank bioreactor based on the design developed using Taguchi L4 method. Results reveal that higher multiplicity of infection (MOI) and size of cell inoculums can yield higher virus titer. Finally, virus samples were purified using high-speed centrifugation based on3∗∗(3-1) Fractional Factorial Design. Statistical analysis showed that the maximum virus titer can be achieved at virus sample concentration of 58.45% (v/v), centrifugation speed of 13729 rpm, and centrifugation time of 4 hours. As a conclusion, high yield of virus titer could be achieved through optimization of cell culture in bioreactor and separation by high-speed centrifugation.

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Mycobacterium abscessus Infection in an Owl Monkey (Aotus trivirgatus)

Pathologia veterinaria ◽

10.1177/030098587000700505 ◽

1970 ◽

Vol 7 (5) ◽

pp. 448-454 ◽

Cited By ~ 1

Author(s):

Alfred G. Karlson ◽

Herman R. Seibold ◽

Robert H. Wolf

Keyword(s):

Cell Culture ◽

Vero Cell ◽

Intraperitoneal Injection ◽

Herpes Virus ◽

Vero Cells ◽

Mycobacterium Abscessus ◽

Vero Cell Culture ◽

Owl Monkey ◽

Herpès Virus

Mycobacterium abscessus was isolated from the lungs of an owl monkey which died 27 days after intraperitoneal injection of herpes virus-infected Vero cells. The lungs and liver had multiple microscopic granulomas with acid-fast microorganisms. The mycobacteria also were isolated from a Vero-cell culture inoculated with a suspension of lung and liver. The same microorganism was eventually isolated from Vero cells of the same source as that used to propagate the herpes virus for the original attempt to infect the monkey.

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