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Cultivation of Bacillus piliformis (Tyzzer) in mouse fibroblasts (3T3 cells)
T H Spencer1, J R Ganaway, K S Waggie
1Comparative Pathology Section, National Institutes of Health, Bethesda, MD 20892.
Abstract:
Bacillus piliformis was successfully propagated in a continuous mouse-embryo fibroblast cell line (3T3). 3T3 monolayers were successfully inoculated with a variety of infected materials including yolk sac and liver suspensions, minced yolk sac and liver, and primary chicken-embryo liver-cell cultures. An initial decrease in B. piliformis number was noted after inoculation of the cell layer with 2.6 X 10(5) organisms followed by a peak bacterial count at 48 h. Bacillus piliformis remained infectious for mice after 22 passages in 3T3 cell monolayers. Limited growth of B. piliformis was obtained in cell lines of mouse connective-tissue origin (L-929) and mouse liver origin (NCTC 1469).
Insights
Bacillus piliformis can be cultured in 3T3 mouse fibroblast cells, showing peak growth at 48 hours. This bacterium remained infectious after multiple passages, indicating a viable method for propagation.
Area of Science:
- Microbiology
- Cell Biology
- Infectious Diseases
Background:
- Bacillus piliformis is an obligate intracellular bacterium.
- Previous studies have focused on its pathogenesis in vivo.
- A reliable in vitro propagation method is needed for further research.
Purpose of the Study:
- To establish and optimize a cell culture system for Bacillus piliformis propagation.
- To assess the viability and infectivity of Bacillus piliformis after in vitro culture.
Main Methods:
- Propagation of Bacillus piliformis in a continuous mouse-embryo fibroblast cell line (3T3).
- Inoculation of 3T3 monolayers using various infected materials.
- Monitoring bacterial growth and infectivity over time and passages.
Main Results:
- Successful propagation of Bacillus piliformis in 3T3 cell monolayers.
- Observed peak bacterial count at 48 hours post-inoculation.
- Bacillus piliformis maintained infectivity for mice after 22 passages in 3T3 cells.
- Limited growth in L-929 and NCTC 1469 cell lines.
Conclusions:
- 3T3 cell line is a suitable host for Bacillus piliformis propagation.
- In vitro cultured Bacillus piliformis retains infectivity, enabling further study.
- Alternative cell lines showed limited success for B. piliformis growth.
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