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Mycobacterium lepraemurium in Cultured Mouse Peritoneal Macrophage

Yonsei Medical Journal 1968년 9권 1호 p.38 ~ 46
양용태 ( Yang Yong-Tae ) - Yonsei University College of Medicine Department of Microbiology

 ( Lew Joon ) - Yonsei University College of Medicine Department of Microbiology

Abstract


Efforts have been made to accomplish a long term in vitro cultivation of mouse peritoneal macrophage as a host cell for growth of Mycobacterium lepraemurium. Following the inoculation with live or heat-killed Myco. lepraemurium of cultured macrophages either on a cover-slip in Leighton tube or in a small petri dish, microscopic observations of acid-fast (AF) stained slide preparation, and total counts of AF bacilli that were released by ultrasonic treatment from the macrophages in small petri dish have been followed in order to present microscopic and quantitative evidence of the actual multiplication of Myco. lepraemurium in cultured mouse peritoneal macrophage. The results are summarized and conclusions are as follows;
1. Successful long term in vitro cultivation of mouse peritoneal macrophage has been accomplished. The growth medium for tissue culture consisted of NCTC 109;50%, heat-inactivated calf serum; 40% and beef embryo extract (diluted 1 : 5) ; 10% and the medium was renewed every 3 to 4 days. The incubation temperature was 37℃ before and at 30℃ after the inoculation with Myco. lepramurium. The CO₂ content inside the CO₂humidity incubator for the cultivation of macrophage was kept at 5%.
2. In cultures of macrophage inoculated with live Myco. lepraemurium, clear features of increases in the number of AF bacilli inside individual cell, of elongation of bacill and of increased solidity in AF staining were observed. However, these features were absent in cultures of macrophage inoculated with heat-killed Myco. lepraemurium.
3. The ultrasonic treatment of macrophage inoculated with live Myco. lepraemurium, and the quantitative assessment of total number of AF bacilli through the course of 6 to 8 weeks after inoculation has provided partial but substantial evidence of actual multiplication of Myco. lepraemurium in cultured macrophages.

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