Characterization of gentamicin-resistant Mycoplasma hyorhinis

Idowu, A. D.; Celones, Michelle C; Salgado, Amparo E; Marquez, Kristine M; Pleibel, Nancy

doi:10.1016/s1045-1056(03)00036-8

Cited by 3 publications

(3 citation statements)

References 9 publications

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“…The incidence of mycoplasma contamination is high and difficult to detect and eliminate (Uphoff et al 1992). Therapeutic products for human or veterinary use require mycoplasma screening at various stages of production, such as during cell banking, virus seed stock preparation, unprocessed bulk harvesting, raw material use, and final product lot release (Idowu et al 2003). According to the US Food and Drug Administration (FDA) and the European Pharmacopoeia, culture and DNA staining methods are the recommended assays for detecting mycoplasma (Eldering et al 2004).…”

Section: Discussionmentioning

confidence: 99%

“…The optimal approach for routine mycoplasma testing is a combination of several techniques. The current industry standard for mycoplasma testing is specified in the US Food and Drug Administration (FDA) guidelines, which involves a culture method and indicator cell culture approach (FDA 1993;Idowu et al 2003). The culture test is performed by inoculating cell samples directly into enriched broth to amplify low-level contaminants for subsequent detection on agar plates.…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, delayed sample analysis is not rare in clinical and laboratory practice, especially when samples are shipped from distant laboratories, when the analysis cannot be performed immediately, or when retesting is required. Good laboratory practices and quality control plans, including sample collection, handling, and storage account for good sample quality and ensure the validity of biological assays (Idowu et al 2003). Some studies have examined the viability of mycoplasma species from humans and other animals (Raccach et al 1975;Furr and Taylor-Robinson, 1990;Zain and Bradbury 1996;Biddle et al 2004).…”

Section: Introductionmentioning

confidence: 99%

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Effect of storage conditions on detection of mycoplasma in biopharmaceutical products

Cheng

Shen

Wang

2007

In Vitro Cell.Dev.Biol.-Animal

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Mycoplasma contamination affects many different aspects of cell culturing, resulting in unreliable experimental results and potentially harmful biological products. Therefore, the specificity, sensitivity, and reliability of detecting mycoplasma contamination are important aspects of quality control in biotechnological products. In this study, Mycoplasma hyorhinis was adopted as a model strain to evaluate the effects of storage on the viability of Mycoplasma species in cell culture samples. Medium X was compared with conventional media 243 and 988 for the ability to detect M. hyorhinis. The 10(1) CFU/ml of M. hyorhinis was inoculated into medium X prepared using the same lots of components and preserved for 7 d, 1 mo, and 2 mo. M. hyorhinis grew readily and typically on agar plates prepared within 1 mo. The viable mycoplasmas in samples containing different initial titers (10(1) and 10(6) CFU/ml) after storage at 4 degrees C and -30 degrees C were analyzed. During storage, viable organisms were found with little or no reduction in titers after storage for 8 wk at -30 degrees C under aerobic and anaerobic conditions. A reduction in titers of 3 log10 occurred after 4 wk storage for high-dose cultures (10(6) CFU/ml) at 4 degrees C. The titers of viable organisms were diminished over 8 wk at 4 degrees C under aerobic and anaerobic conditions.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effect of storage conditions on detection of mycoplasma in biopharmaceutical products

Cheng

Shen

Wang

2007

In Vitro Cell.Dev.Biol.-Animal

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Development of a PCR method for mycoplasma testing of Chinese hamster ovary cell cultures used in the manufacture of recombinant therapeutic proteins

Eldering¹,

Felten²,

Veilleux³

et al. 2004

Biologicals

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Impact of Mycoplasma hyorhinis infection on l-arginine metabolism: differential regulation of the human and murine iNOS gene

Kagemann

Henrich

Kühn

et al. 2005

Biological Chemistry

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Infection with mycoplasma is a common problem in cell cultures, with Mycoplasma hyorhinis being the predominant species. Here we investigate the effect of M. hyorhinis infection on L-arginine metabolism, with focus on iNOS-mediated NO synthesis in murine keratinocytes and the human colon cancer cell line DLD-1. iNOS and arginase are L-arginine-metabolizing enzymes involved in the regulation of inflammatory processes, with NO contributing to innate immunity. In murine cells, M. hyorhinis infection enhances cytokine-induced iNOS expression and augments iNOS activity, whereas in the absence of cytokines it causes de novo induction of iNOS mRNA without subsequent translation into iNOS protein. In turn, arginase-1 mRNA expression is diminished in M. hyorhinis-infected murine keratinocytes, resulting in decreased arginase activity. One of the underlying upstream mechanisms is NF-kappaB activation. In contrast, in human cells neither iNOS mRNA nor protein expression is affected by M. hyorhinis infection, but NO synthesis is enhanced, which may be caused by increased L-arginine import. This demonstrates that infection with M. hyorhinis leads to different effects on gene regulation of the murine and human iNOS gene. Our study underlines the importance of routine checking of cell cultures for mycoplasma contamination, particularly in studies on NO-mediated effects or inflammatory processes.

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Characterization of gentamicin-resistant Mycoplasma hyorhinis

Cited by 3 publications

References 9 publications

Effect of storage conditions on detection of mycoplasma in biopharmaceutical products

Effect of storage conditions on detection of mycoplasma in biopharmaceutical products

Development of a PCR method for mycoplasma testing of Chinese hamster ovary cell cultures used in the manufacture of recombinant therapeutic proteins

Impact of Mycoplasma hyorhinis infection on l-arginine metabolism: differential regulation of the human and murine iNOS gene

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