Growth of Mycobacterium leprae in Macrophages in vitro

概要

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Macrophages were obtained from the peritoneal cavities of mice and infected in vitro with Mycobacterium leprae. After three weeks, the multiplication of the bacteria was observed inside of the infected cells. Three weeks after the infection the medium was replaced and antibiotic free medium was added to culture flask which was incubated at 32°C. Three months later, amorphous turbidity was observed under the surface layer of the medium. This turbidity slowly progressed with time. Next the sample was centrifuged and CBA macrophages ingested. After 3 weeks Ziehl-Neelsen positive rods were observed in 0.2-0.6% of the cells. These observations not only indicated the release of M. leprae from infected macrophages, but also suggested the possibility of the micro-organism growing in the medium itself. With these observations and experiences accumulated over fifteen years of trial and error, I discuss various potential improvements for the medium and the environment to permit more efficient growth of M. leprae in vitro.
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