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A microcarrier culture method for the production of large quantities of viable Chlamydia pneumoniae
A Meijer1, C E Vallinga, J M Ossewaarde
1National Institute of Public Health and The Environment, Department of Virology, Bilthoven, The Netherlands.
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 method using centrifugation-assisted infection. In conclusion, the microcarrier culture method is useful to produce large quantities of viable Chlamydia pneumoniae economically.
Insights
Microcarrier culture effectively propagates Chlamydia pneumoniae, offering significant labor and cost savings compared to traditional methods. This technique provides an economical way to produce large quantities of viable bacteria.
Area of Science:
- Microbiology
- Cell Culture Technology
Background:
- Chlamydia pneumoniae is a significant human pathogen.
- Efficient large-scale culture of C. pneumoniae is crucial for research and diagnostics.
Purpose of the Study:
- To optimize and evaluate the microcarrier bead culture method for propagating Chlamydia pneumoniae.
- To compare the economic viability and efficiency of microcarrier culture versus traditional microtitre-plate culture.
Main Methods:
- HEp2 cells were cultured on microcarrier beads.
- Infection was optimized using polyethylene glycol 4000 (PEG4000) during adsorption.
- Bacterial yield and burst size were compared between microcarrier and microtitre-plate cultures.
Main Results:
- Microcarrier culture yielded comparable amounts of viable Chlamydia pneumoniae to microtitre-plate culture.
- Significant savings in labor (up to 64%) and culture medium (27%) were achieved with microcarrier culture.
- Optimal bacterial yield was independent of infection mode and incubation temperature (35°C or 37°C) in microcarrier cultures.
Conclusions:
- Microcarrier bead culture is an efficient and economical method for large-scale production of viable Chlamydia pneumoniae.
- This method offers substantial cost and labor benefits for researchers and diagnostic laboratories.
