TY - JOUR
T1 - Evaluation of the cell growth of mycobacteria using Mycobacterium smegmatis mc2 155 as a representative species
AU - Gonzalez-y-Merchand, Jorge A.
AU - Zaragoza-Contreras, Ruben
AU - Guadarrama-Medina, Rosalina
AU - Helguera-Repetto, Addy C.
AU - Rivera-Gutierrez, Sandra
AU - Cerna-Cortes, Jorge F.
AU - Santos-Argumedo, Leopoldo
AU - Cox, Robert A.
N1 - Funding Information:
We thank Simon A. Cox for his help in the preparation of this manuscript, Iris Estrada-Garcia for helpful review of the manuscript, and Oliver Ornelas-Lopez and Esther Ramirez-Espindola from the Microscopy Unit of ENCB, IPN, for their help with the electron microscopy analysis. This work is supported as part of the 7th Framework Program, European Commission, contract no. HEALTH-F3-2008-200999. It was also partially supported by grants SIP20110750, SIP-20110106 and SIP 20110448 from IPN, Mexico. J.A.G-y-M, J.F.C-C, S.R-G and A.C.H-R are fellows of COFAA and EDI, IPN, Mexico.
PY - 2012/6
Y1 - 2012/6
N2 - 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.
AB - 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.
KW - Mycobacterium smegmatis
KW - flow cytometry
KW - mycobacterial growth
KW - mycobacterial physiology
UR - http://www.scopus.com/inward/record.url?scp=84863318999&partnerID=8YFLogxK
U2 - 10.1007/s12275-012-1556-0
DO - 10.1007/s12275-012-1556-0
M3 - Artículo
C2 - 22752905
SN - 1225-8873
VL - 50
SP - 419
EP - 425
JO - Journal of Microbiology
JF - Journal of Microbiology
IS - 3
ER -