A microcarrier culture method for the production of large quantities of viable Chlamydia pneumoniae

Applied Microbiology and Biotechnology
A MeijerJ M Ossewaarde

Abstract

We studied the propagation of Chlamydia pneumoniae strain TW-183 in HEp2 cells grown on microcarrier beads. Infection of the cells in microcarrier culture was optimized by addition of 7.5% polyethylene glycol 4000 (PEG4000) during adsorption. The yield in microcarrier culture was similar to that of microtitre-plate culture using centrifugation-assisted infection (120 x 10(6) and 225 x 10(6) bacteria/10(6) HEp2 cells respectively), as was the burst size (505 and 449 bacteria produced/infecting bacterium respectively). However, up to 64% savings in labour time and 27% saving in culture medium were achieved if the microcarrier culture method was used instead of the microtitre-plate culture method. The optimal yield of viable bacteria could only be achieved at a narrow range of multiplicities of infection (0.24 - 1.14 inclusion-forming units/cell), independent of the mode of infection (centrifugation-assisted infection of PEG4000-facilitated infection by adsorption) and independent of incubation temperature (35 degrees C or 37 degrees C). The yield of microcarrier cultures was the same at an incubation temperature of 35 degrees C or 37 degrees C in contrast to an increased production at 35 degrees C in the microtitre-plate culture ...Continue Reading

Citations

Jun 15, 2004·International Journal of Cardiology·Gülay YetkinAli Mert
Mar 27, 2001·Journal of Clinical Pathology·A MeijerJ M Ossewaarde
Jan 29, 2005·Journal of Microbiological Methods·Dan LiBernhard Kaltenboeck
Nov 1, 2005·Vaccine·Isabel MarcelinoPaula M Alves
Mar 30, 2006·Biotechnology and Bioengineering·P van HeeL A M van der Wielen
Mar 13, 1999·Journal of Clinical Microbiology·A TiranM M Wilders-Truschnig

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