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Growth of Leptospira pomona and Its Effect on Various Tissue Culture Systems
R E Miller1, N G Miller, R J White
1Department of Medical Microbiology, University of Nebraska College of Medicine, Omaha, Nebraska.
Abstract:
Miller, Robert E. (University of Nebraska College of Medicine, Omaha), Norman G. Miller, and Roberta J. White. Growth of Leptospira pomona and its effect on various tissue culture systems. J. Bacteriol. 92:502-509. 1966.-Leptospira pomona strain 3341 was grown in association with primary fetal bovine kidney (PBK) and human embryonic skin-muscle fibroblastic (HE) cells in Eagle's minimal essential medium (MEM) with 5% sheep serum. Growth curves of leptospires in PBK and HE cell cultures showed no substantial increase in growth above that obtained in Eagle's MEM in the absence of tissue culture cells. This suggested that no stimulatory growth factors for leptospires were produced by the tissue cells. Fibroblastic cells of the PBK monolayer showed separation, deterioration, and, finally, complete disintegration. Epithelial-like cells remained unaffected. HE cells showed the same cytopathic effect as PBK fibroblastic cells, indicating that this effect was not limited to PBK fibroblastic cells. Warthin-Starry stains of PBK and HE cell monolayers showed masses of leptospires adhering to fibroblastic cells, whereas only a few were seen on epithelial-like cells. Large numbers of leptospires on the surface of fibroblastic cells are very likely associated with the cytopathic effect. Dislodgment of leptospires from fibroblastic cells did not increase the total number of spirochetes in the culture. This indicated that leptospiral growth did not occur on the surface of these cells.
Insights
Leptospira pomona did not grow significantly in tissue culture systems. The bacteria caused damage to fibroblastic cells, but not epithelial cells, suggesting a specific interaction.
Area of Science:
- Microbiology
- Cell Biology
- Bacteriology
Background:
- Leptospira pomona is a bacterial spirochete.
- Understanding bacterial growth and host cell interactions is crucial in microbiology.
Purpose of the Study:
- To investigate the growth of Leptospira pomona in association with tissue culture systems.
- To determine the effects of Leptospira pomona on different cell types within these cultures.
Main Methods:
- Leptospira pomona strain 3341 was cultured with primary fetal bovine kidney (PBK) and human embryonic skin-muscle fibroblastic (HE) cells.
- Growth curves were monitored in Eagle's minimal essential medium (MEM) with sheep serum.
- Warthin-Starry stains were used to visualize leptospires on cell monolayers.
Main Results:
- Leptospira pomona showed no significant growth increase when cultured with PBK or HE cells compared to cell-free medium.
- Fibroblastic cells (PBK and HE) exhibited deterioration and disintegration, indicating a cytopathic effect.
- Leptospires were observed adhering predominantly to fibroblastic cells, with minimal association with epithelial-like cells.
Conclusions:
- Tissue culture cells do not provide stimulatory growth factors for Leptospira pomona.
- The cytopathic effect on fibroblastic cells is likely due to the large numbers of leptospires adhering to their surface.
- Leptospiral growth does not appear to occur on the surface of these tissue culture cells.
