The relevance of antibiotic supplements in mammalian cell cultures: Towards a paradigm shift

Hassan, Siti Nazihahasma; Ahmad, Fadzil

doi:10.4274/gulhane.galenos.2020.871

Cited by 10 publications

(6 citation statements)

References 28 publications

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“…In addition to contaminant importation, intracellular variation in some indigenous particles can be mistaken for bacteria at times 13,14 . Other intracellular organism of unclear nature which may have a symbiotic association with cell line has also been linked to cell line contamination 6 . Despite black dot contamination, cell growth and morphology were healthy.…”

Section: Discussionmentioning

confidence: 99%

“…Viral and mycoplasma contamination do not cause noticeable turbidity in the media, have a lower pH 4,5 , and are better tolerated by cells. They are only excluded from cell lines if they are periodically checked and subjected to suitable processing and modification requirements 6 . The black spots in the cultured cells are remarkably little dots.…”

Section: Introductionmentioning

confidence: 99%

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Elucidating the Effectiveness of Antibiotic Treatments in Eliminating the Black Dots Contamination from Vero Cell Line Culture

Barkathullah¹,

Sonowal²,

Mohd³

et al. 2022

J. Pure Appl. Microbiol.

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The present study was conducted to identify the effectiveness of various antibiotics and antibiotic combinations in eliminating the black dots contamination in cell culture and to identify its effect on cell growth kinetics. Isolation of contaminant organism (Black dots) observed in our laboratory, was attempted by inoculation of harvested cell supernatant and pellet into Nutrient broth, Luria broth, and Pleuropneumonia-like organisms’ broth but no visible growth was observed. To analyze the effect of antibiotics in elimination of contamination, the cell monolayer was incubated with different antibiotics and combination of antibiotics till 120 hrs. Withdrawal effect of antibiotics was observed by further incubation cells in maintenance media for another 48 hrs. To investigate the effect of different antibiotics on cell growth kinetics, cell seeding was done in media containing antibiotics. A subset of cells was harvested every 24 hrs and counted till 120 hrs. Among all antibiotics, Ampicillin reduced the contamination, the Piperacillin and Tazobactam combination diminished the contamination to a great extent. At the studied concentrations of the antibiotic trio of piperacillin, tazobactam, and ampicillin, black dot contamination was eliminated and found 100% effective in inhibiting contamination within 120 hrs of the study period; but, after 48 hrs of antibiotic withdrawal, contamination reappears in cell culture. The growth kinetics showed no significant impact of antibiotics treatment on the cell growth curve. The growth kinetics showed that cell growth was not interfering significantly by individual or combination of antibiotics. Therefore, in the presence of black dot, a combination of three antibiotics (piperacillin, tazobactam, and ampicillin) should be effective in cell culture.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Elucidating the Effectiveness of Antibiotic Treatments in Eliminating the Black Dots Contamination from Vero Cell Line Culture

Barkathullah¹,

Sonowal²,

Mohd³

et al. 2022

J. Pure Appl. Microbiol.

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“…Immortalized cell lines were employed to avoid impacts associated with primary cell senescence and repeated chemical risk assessments. , Antibiotics were not used in the medium as they can alter gene expression and have cytotoxic and cytostatic activities, which could result in inaccurate results. , …”

Section: Materials and Methodsmentioning

confidence: 99%

Bioenergetic Shifts in Humpback Whale Fibroblasts Upon Chemical Exposure

Hosen,

Whitworth,

Leusch

et al. 2024

Environ. Sci. Technol.

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Southern Hemisphere humpback whales accumulate persistent and toxic chemicals, which are transported to Antarctica through distant sources and in situ usage. The extreme seasonal migration-associated fast of humpback whales results in the remobilization of persistent and lipophilic environmental contaminants from liberated fat stores. Mitochondria play a key role in lipid metabolism, and any disruption to mitochondrial function is expected to influence whole-organism bioenergetics. It is therefore of interest to advance understanding of the impact of known contaminants of the Antarctic sea-ice ecosystem upon humpback whale cellular bioenergetics. Using cell line-based in vitro testing, this study employed the Seahorse Extracellular Flux Analyzer to study cellular metabolic activity in live humpback whale fibroblast cells. The assay, based on oxygen consumption rate, provides insights into the cause of cellular bioenergetic disruption. Immortalized skin fibroblasts were exposed to four priority environmental chemicals found in the Antarctic sea-ice ecosystem. Our findings reveal chemical-dependent functional alterations and varying bioenergetic profile responses. Chlorpyrifos was observed to decrease mitochondrial basal oxygen consumption; dieldrin increased basal oxygen consumption; trifluralin's impact was dose-specific, and endosulfan displayed no effect. Our results provide unique insights into environmental chemical mechanisms of action on cellular bioenergetics, generating much-needed taxa-specific chemical effect data in support of evidence-based conservation policy and management.

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“…Antibiotics were not used in the medium as they can alter gene expression, as well as have cytotoxic and cytostatic activity, which could result in inaccurate results. , …”

Section: Materials and Methodsmentioning

confidence: 99%

Novel Use of Cell Profiling Technology to Visualize Mitochondrial Responses of Humpback Whale Fibroblasts to Chemical Exposure

Hosen

Sykes

Wood

et al. 2023

Environ. Sci. Technol.

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Cetaceans are at elevated risk of accumulating persistent and lipophilic environmental contaminants due to their longevity and high proportion of body fat. Despite this, there is a paucity of taxa-specific chemical effect data, in part due to the ethical and logistical constraints in working with highly mobile aquatic species. Advances in cetacean cell culture have opened the door to the application of mainstream in vitro toxicological effect assessment approaches. Image-based cell profiling is a high-throughput, microscopy-based system commonly applied in drug development. It permits the analysis of the xenobiotic effect on multiple cell organelles simultaneously, hereby flagging its potential utility in the evaluation of chemical toxicodynamics. Here we exposed immortalized humpback whale skin fibroblasts (HuWaTERT) to six priority environmental contaminants known to accumulate in the Southern Ocean food web, in order to explore their subcellular organelle responses. Results revealed chemical-dependent modulation of mitochondrial texture, with the lowest observed effect concentrations for chlorpyrifos, dieldrin, trifluralin, and p,p’-dichlorodiphenyldichloroethane of 0.3, 4.1, 9.3, and 19.8 nM, respectively. By contrast, no significant changes were observed upon exposure to endosulfan and lindane. This study contributes the first fixed mitochondrial images of HuWaTERT and constitutes novel, taxa-specific chemical effect data in support of evidence-based conservation policy and management.

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The relevance of antibiotic supplements in mammalian cell cultures: Towards a paradigm shift

Cited by 10 publications

References 28 publications

Elucidating the Effectiveness of Antibiotic Treatments in Eliminating the Black Dots Contamination from Vero Cell Line Culture

Elucidating the Effectiveness of Antibiotic Treatments in Eliminating the Black Dots Contamination from Vero Cell Line Culture

Bioenergetic Shifts in Humpback Whale Fibroblasts Upon Chemical Exposure

Novel Use of Cell Profiling Technology to Visualize Mitochondrial Responses of Humpback Whale Fibroblasts to Chemical Exposure

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