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An improved method for the prolonged maintenance of Pneumocystis carinii in vitro
1Infectious Disease Unit, Massachusetts General Hospital, Boston.
Abstract:
A culture system for rat-derived Pneumocystis carinii was developed that allows the long-term maintenance of viable organisms in vitro. Organisms were derived from the lungs of immunosuppressed rats infected with P. carinii. P. carinii was maintained in culture with human embryonal lung (HEL) fibroblasts and Eagle MEM for up to 42 days. Passage of organisms was done by reinoculation of infected HEL cells with attached P. carinii onto a fresh monolayer of confluent cells. Rats pretreated with trimethoprim-sulfamethoxazole and inoculated with cultured organisms (up to day 42) developed P. carinii pneumonia by 6 weeks while control animals remained uninfected. P. carinii was able to be harvested from these lungs and reintroduced into tissue culture. Cultures have been maintained for up to 6 weeks without a substantial loss of P. carinii viability. In combination with intratracheal inoculation, this system raises the possibility of maintaining stable laboratory strains of P. carinii.
Insights
A new in vitro culture system enables long-term maintenance of viable Pneumocystis carinii, facilitating research into this opportunistic pathogen. This breakthrough allows for the development of stable laboratory strains for further study.
Area of Science:
- Microbiology
- Infectious Diseases
- Cell Culture
Background:
- Pneumocystis carinii pneumonia (PCP) is a significant opportunistic infection.
- Maintaining viable Pneumocystis carinii organisms in vitro has been a challenge for research.
- Previous methods limited the study of P. carinii lifecycle and pathogenesis.
Purpose of the Study:
- To develop a reliable in vitro culture system for rat-derived Pneumocystis carinii.
- To enable long-term maintenance of viable Pneumocystis carinii organisms.
- To establish a foundation for creating stable laboratory strains of P. carinii.
Main Methods:
- Rat-derived Pneumocystis carinii were cultured using human embryonal lung (HEL) fibroblasts and Eagle MEM.
- Organisms were maintained in culture for up to 42 days.
- Passage was achieved by reinoculating fresh HEL cell monolayers with infected cells.
Main Results:
- Viable Pneumocystis carinii were maintained in culture for up to 42 days.
- Inoculation of rats with cultured organisms (up to day 42) successfully induced Pneumocystis carinii pneumonia.
- Organisms could be harvested from infected rat lungs and reintroduced into tissue culture, maintaining viability for up to 6 weeks.
Conclusions:
- A robust in vitro culture system for rat-derived Pneumocystis carinii has been established.
- This system supports long-term viability and infectivity of Pneumocystis carinii.
- The developed culture method offers a promising approach for maintaining stable laboratory strains for research.