Nutritional effects of culture media on mycoplasma cell size and removal by filtration

Folmsbee, Martha; Howard, G. A.; McAlister, Morven

doi:10.1016/j.biologicals.2009.11.001

Cited by 28 publications

(14 citation statements)

References 25 publications

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“…Members of the Acholeplasma genus do not have specific host associations and have been found in almost every type of living organism. They are one of the five common reasons of cell culture contamination, and a causative agent of some plant diseases [44], [45], [46], [47].…”

Section: Discussionmentioning

confidence: 99%

Metabolomic Analysis of Three Mollicute Species

et al. 2014

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We present a systematic study of three bacterial species that belong to the class Mollicutes, the smallest and simplest bacteria, Spiroplasma melliferum, Mycoplasma gallisepticum, and Acholeplasma laidlawii. To understand the difference in the basic principles of metabolism regulation and adaptation to environmental conditions in the three species, we analyzed the metabolome of these bacteria. Metabolic pathways were reconstructed using the proteogenomic annotation data provided by our lab. The results of metabolome, proteome and genome profiling suggest a fundamental difference in the adaptation of the three closely related Mollicute species to stress conditions. As the transaldolase is not annotated in Mollicutes, we propose variants of the pentose phosphate pathway catalyzed by annotated enzymes for three species. For metabolite detection we employed high performance liquid chromatography coupled with mass spectrometry. We used liquid chromatography method - hydrophilic interaction chromatography with silica column - as it effectively separates highly polar cellular metabolites prior to their detection by mass spectrometer.

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Section: Discussionmentioning

confidence: 99%

Metabolomic Analysis of Three Mollicute Species

et al. 2014

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“…About 700 protein spots were resolved in the analytic gel. Probably, the low resolution is connected with more broad pH range of IPG we used (3-10) then it is mainly used in the plant proteomic study (3)(4)(5)(6)(7).…”

Section: Resultsmentioning

confidence: 99%

“…This mycoplasma is widely distributed in nature, one of the five common species of cell culture contaminants, and is a causative agent of some plant diseases [3][4][5][6][7]. Insects are that the adaptation of A. laidlawii PG8 to starvation involves changes in morphology, ultrastructure, DNA topology, genotoxicity of the mycoplasma cells and virulence of the bacterium toward Vinca minor L. -the specific indicator of phytomycoplasmoses.…”

Section: Introductionmentioning

confidence: 99%

Unadapted and adapted to starvation Acholeplasma laidlawii cells induce different responses of Oryza sativa, as determined by proteome analysis

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Medvedeva

et al. 2011

Journal of Proteomics

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“…Mycoplasmas infect the cell cultures so easily and may not be visible and detectable by ordinary optical microscopic tools. Their flexibility, small-scale and polymorphism allows mycoplasmas to easily cross through anti-bacterial filters with a diameter of 0.45-0.22 lm (Folmsbee et al 2010;Waites et al 2013). Studies indicate that up to 87 % of cell lines in different cell banks are infected with mycoplasmas.…”

Section: Introductionmentioning

confidence: 99%

Real-time PCR assay is superior to other methods for the detection of mycoplasma contamination in the cell lines of the National Cell Bank of Iran

et al. 2015

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Mycoplasmas are the most important contaminants of cell cultures throughout the world. They are considered as a major problem in biological studies and biopharmaceutical economic issues. In this study, our aim was to find the best standard technique as a rapid method with high sensitivity, specificity and accuracy for the detection of mycoplasma contamination in the cell lines of the National Cell Bank of Iran. Thirty cell lines suspected to mycoplasma contamination were evaluated by five different techniques including microbial culture, indirect DNA DAPI staining, enzymatic mycoalert Ò assay, conventional PCR and real-time PCR. Five mycoplasma-contaminated cell lines were assigned as positive controls and five mycoplasma-free cell lines as negative controls. The enzymatic method was performed using the mycoalert Ò mycoplasma detection kit. Real-time PCR technique was conducted by PromoKine diagnostic kits. In the conventional PCR method, mycoplasma genus-specific primers were designed to analyze the sequences based on a fixed and common region on 16S ribosomal RNA with PCR product size of 425 bp. Mycoplasma contamination was observed in 60, 56.66, 53.33, 46.66 and 33.33 % of 30 different cell cultures by real-time PCR, PCR, enzymatic mycoalert Ò , indirect DNA DAPI staining and microbial culture methods, respectively. The analysis of the results of the different methods showed that the realtime PCR assay was superior the other methods with the sensitivity, specificity, accuracy, predictive value of positive and negative results of 100 %. These values were 94.44, 100, 96.77, 100 and 92.85 % for the conventional PCR method, respectively. Therefore, this study showed that real-time PCR and PCR assays based on the common sequences in the 16S ribosomal RNA are reliable methods with high sensitivity, specificity and accuracy for detection of mycoplasma contamination in cell cultures and other biological products.

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Nutritional effects of culture media on mycoplasma cell size and removal by filtration

Cited by 28 publications

References 25 publications

Metabolomic Analysis of Three Mollicute Species

Metabolomic Analysis of Three Mollicute Species

Unadapted and adapted to starvation Acholeplasma laidlawii cells induce different responses of Oryza sativa, as determined by proteome analysis

Real-time PCR assay is superior to other methods for the detection of mycoplasma contamination in the cell lines of the National Cell Bank of Iran

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