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Evaluation of the cell growth of mycobacteria using Mycobacterium smegmatis mc2 155 as a representative species

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Abstract

The study of the in vitro cell growth of mycobacteria still remains a fastidious, difficult, and time-consuming procedure. In addition, assessing mycobacterial growth in the laboratory is often complicated by cell aggregation and slow growth-rate. We now report that the use of a stainless steel spring in the culture led to an absence of large cell clumps, to a decrease of dead cells in the exponential phase and to growth of a more homogeneous population of large cells. We also report that flow cytometry is a rapid, simple and reliable approach to monitor mycobacterial cell growth and viability. Here, we monitored Mycobacterium smegmatis cellular growth by optical density, dry cell mass, and colony forming units; in addition, viability, cell size and granularity profiles were analyzed by flow cytometry, and cell morphology by electron microscopy. Cultures monitored by flow cytometry may lead to a better understanding of the physiology of mycobacteria. Moreover, this methodology may aid in characterizing the cell growth of other fastidious species of microorganisms.

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Authors and Affiliations

  1. Departamento de Microbiologia, Escuela Nacional de Ciencias Biologicas, IPN, Prolong. Carpio y Plan de Ayala s/n, Col. Casco de Santo Tomas, Mexico, D.F., 11340, Mexico

    Jorge A. Gonzalez-y-Merchand, Ruben Zaragoza-Contreras, Rosalina Guadarrama-Medina, Addy C. Helguera-Repetto, Sandra Rivera-Gutierrez & Jorge F. Cerna-Cortes

  2. Departamento de Biomedicina Molecular, CINVESTAV, Zacatenco, Mexico, D.F., Mexico

    Leopoldo Santos-Argumedo

  3. Division of Mycobacterial Research, National Institute for Medical Research, The Ridgeway, Mill Hill, London, NW7 1AA, UK

    Robert A. Cox

Authors
  1. Jorge A. Gonzalez-y-Merchand
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  2. Ruben Zaragoza-Contreras
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  3. Rosalina Guadarrama-Medina
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  4. Addy C. Helguera-Repetto
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  5. Sandra Rivera-Gutierrez
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  6. Jorge F. Cerna-Cortes
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  7. Leopoldo Santos-Argumedo
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  8. Robert A. Cox
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Correspondence to Jorge A. Gonzalez-y-Merchand.

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Gonzalez-y-Merchand, J.A., Zaragoza-Contreras, R., Guadarrama-Medina, R. et al. Evaluation of the cell growth of mycobacteria using Mycobacterium smegmatis mc2 155 as a representative species. J Microbiol. 50, 419–425 (2012). https://doi.org/10.1007/s12275-012-1556-0

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  • DOI: https://doi.org/10.1007/s12275-012-1556-0

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