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Cell Culture Contaminants Identification

Cell culture is an important in vitro method by which cells, usually extracted from plants or animals, are grown in the lab. One of the major challenges in cell culture is to properly manage contamination. The contaminants can be categorized into two forms: chemical contaminants such as impurities in media and water, endotoxins, detergent; and biological contaminants such as bacteria, yeast, viruses, mycoplasma and other cell lines. Among these contaminants, mycoplasma, a genus of bacteria that lack a cell wall, draws particular concerns since they are resistant to many antibiotics such as penicillin. In addition, the absence of a cell wall makes mycoplasma invisible under phase contrast microscopy. As the applications of cell culture increase in research, industrial production and cell therapy, substantial effort has been invested for contamination detection and identification in cell culture. Methods of detecting mycoplasmas contain PCR, DNA staining by fluorochromes, fluorescence in situ hybridization (FISH) and Fourier transform infrared (FTIR) microspectroscopy. These methods provide unmatched versatility and efficacy in detecting biological contamination, but the lack of resolution at molecular level has prevented their application in chemical contaminant detection.

Scanning beam electron microscopy of Mycoplasma. pneumoniae. 1. M. pneumoniae cells during phase I growth (8 hr). Characteristic tightly packed round forms are seen.2. M. pneumoniae during early phase IH growth (2 days). Spherical cells and short, straight, branching filaments are shown. 3. M. pneumoniae during phase II growth (3 days). The characteristic microcolony is composed of intertwined filaments and occasional round forms. 4. M. pneumoniae during mid-phase II growth (4 days). This microcolony is composed of both round and filamentous forms with bulbous elements. 5. M. pneumoniae during the late phase 11 growth (6 days). Shorter filaments with both terminal and intervening bulbs are predominant. 6. M. pneumoniae during phase III growth (10 days). Larger, asymmetrical round forms are shown. Flat, collapsed. Figure 1. Scanning beam electron microscopy of M. pneumoniae.

On the other hand, electron microscope (EM) is a powerful tool to reveal the structural details of biological, organic and inorganic molecules at much higher resolution than optical microscope. At Creative Biostructure, we are devoted to the development of EM strategies and applications in contaminant testing. Combined with negative staining technique, EM can quickly and rapidly detect all cell contaminants, regardless of the molecular size.

Creative Biostructure has the most up-to-date EM platform and a wealth of experience in EM-assisted studies. Our knowledge and capabilities enable us to provide standard and customized services to meet our customer’s requirements. Creative Biostructure promises to work closely with our customers, and we are confident we can find solutions to even the most challenging problems.

Please feel free to contact us for a detailed quote.

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References

  1. Laleh Nikfarjam, Parvaneh Farzaneh, Prevention and Detection of Mycoplasma Contamination in Cell Culture. Cell J. 2012; 13: 203-212.
  2. Gary M. Kammer, J. Dennis Pollack, et al. Scanning-Beam Electron Microscopy of Mycoplasma pneumoniae. J Bacteriol. 1970; 104: 499-502.

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