Detection of &lt;i&gt;Mycoplasma &lt;/i&gt;species in cell culture by PCR and RFLP based method: Effect of BM-cyclin to cure infections

Gopalkrishna, Varanasi; Verma, Harsha; Kumbhar, NS; Tomar, RS; Patil, PR

doi:10.4103/0255-0857.37340

Cited by 9 publications

(4 citation statements)

References 18 publications

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“…in nearly 86% cases. Similar results in the effectivity were achieved with sparfloxacin and BM-Cyclin for decontamination of cell lines (Drexler et al, 1995;Fleckenstein and Drexler, 1996;Drexler and Uphoff, 2002;Gopalkrishna et al, 2007). The problem of cytotoxicity appeared in our case as unimportant.…”

Section: Discussionsupporting

confidence: 84%

Elimination of mycoplasma contamination of virus stocks

Malenovská¹,

Reichelova²

2011

Vet. Med. - Czech

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ABSTRACT:We studied the effectivity of a commercial antibiotic kit Mycokill AB for the elimination of mycoplasma contamination in virus strains. The contaminated virus strains were first filtered, treated with Mycokill AB for three hours and repeatedly passaged in its presence in the cultivation medium in pure cell lines. Three passages in the presence of Mycokill AB were invariably followed by three passages without Mycokill AB. The effectivity of purification was then checked by PCR. Twenty-four out of 28 tested virus strains became free of mycoplasma after a first or second cycle of the treatment with Mycokill AB. The other four strains remained positive even after repeated passages. In these cases of a likely resistance to Mycokill AB, we managed to eliminate the mycoplasma contamination through a subsequent treatment with the antibiotic combination BM-Cyclin. Mycokill AB was shown in the elimination of mycoplasma from virus suspensions as successful as other known most effective antibiotics.

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Section: Discussionsupporting

confidence: 84%

Elimination of mycoplasma contamination of virus stocks

Malenovská¹,

Reichelova²

2011

Vet. Med. - Czech

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“…Cell line characterization was performed by short tandem repeat analysis using GeneMapper ® software (version 4.0; ) and consolidated using the American Type Culture Collection, German Collection of Microorganisms and Cell Cultures, Japanese Collection of Research Bioresources Cell Bank and Rikagaku Kenkyusho (Institute of Physical and Chemical Research, Japan) databases (; ; ). The cell lines were tested for the presence of Mycoplasma using PCR (32).…”

Section: Methodsmentioning

confidence: 99%

SOX9 promotes epithelial‑mesenchymal transition via the Hippo‑YAP signaling pathway in gastric carcinoma cells

Zhou¹,

Li²,

Huang³

et al. 2019

Oncol Lett

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SRY-box 9 (SOX9) is overexpressed in a number of human tumors, including gastric cancer (GC). However, the function of SOX9 in the development of GC remains unknown. In the present study, SOX9 activated the Hippo-yes-associated protein (YAP) signaling pathway to enhance the epithelial-mesenchymal transition in GC cell lines. The results suggested that SOX9 knockdown inhibited invasion, proliferation and migration of GC cells. Furthermore, SOX9 silencing upregulated the expression of E-cadherin, an epithelial marker, and downregulated the expression of mesenchymal markers, including snail family transcriptional repressor 1, vimentin and N-cadherin. SOX9 overexpression increased the expression of the aforementioned markers. SOX9 significantly affected YAP phosphorylation and total YAP protein levels, suggesting that SOX9 is involved in the Hippo-YAP signaling pathway. The current study revealed that SOX9 may be involved in the pathogenesis of GC, and further elucidation of the pathways involved may support the development of novel therapeutic options for the treatment of GC.

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“…Cells do not die because of mycoplasma but their metabolism can be altered leading to chromosomal aberrations, decrease in the growth rate and detachment of cells from culture surface. Mycoplasma detection methods are based on commercial kits utilizing PCR reaction and DAPI or Hoechst staining ( Gopalkrishna et al, 2007 ; Armstrong et al, 2010 ; Philippeos et al, 2012 ; Uphoff et al, 2012 ; Faison et al, 2020 ). It is very easy to infect cells by carrying mycoplasma bacteria on our skin, with infected reagents (e.g., serum) or other infected cell culture, thus Good Laboratory Practice (GLP) should be introduced in every experimental laboratory carrying cell culture work.…”

Section: Discussionmentioning

confidence: 99%

An Overview of Methods and Tools for Transfection of Eukaryotic Cells in vitro

Fus-Kujawa

Prus

Bajdak-Rusinek

et al. 2021

Front. Bioeng. Biotechnol.

107

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Transfection is a powerful analytical tool enabling studies of gene products and functions in eukaryotic cells. Successful delivery of genetic material into cells depends on DNA quantity and quality, incubation time and ratio of transfection reagent to DNA, the origin, type and the passage of transfected cells, and the presence or absence of serum in the cell culture. So far a number of transfection methods that use viruses, non-viral particles or physical factors as the nucleic acids carriers have been developed. Among non-viral carriers, the cationic polymers are proposed as the most attractive ones due to the possibility of their chemical structure modification, low toxicity and immunogenicity. In this review the delivery systems as well as physical, biological and chemical methods used for eukaryotic cells transfection are described and discussed.

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Detection of <i>Mycoplasma </i>species in cell culture by PCR and RFLP based method: Effect of BM-cyclin to cure infections

Cited by 9 publications

References 18 publications

Elimination of mycoplasma contamination of virus stocks

Elimination of mycoplasma contamination of virus stocks

SOX9 promotes epithelial‑mesenchymal transition via the Hippo‑YAP signaling pathway in gastric carcinoma cells

An Overview of Methods and Tools for Transfection of Eukaryotic Cells in vitro

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