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KMID : 0386319680050010109
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Korean Leprosy Bulletin
1968 Volume.5 No. 1 p.109 ~ p.115
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Mycobacterium lepraemurium I cultured mouse peritoneal macrophage
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梁容泰/YANG, Y.T
柳駿/韓聖鉉/金昇燁/Lew, J/Han, S.H/Kim, S.Y
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Abstract
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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 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 evidences of the actual multiplication of Myco. lepraemurium in cultured mouse peritoneal macrophage. The results are summarized and concluded as follows;
1. Successful long term in vitro cultivation of mouse peritoneal macrophage has been accomplished. The growth media for tissue culture is consisted of NCTC 109;50 %, heat inactivated calf serum; 40Y. and beef embryo extract (diluted 1 : 5) 10% and the media was renewed every 3 to 4 days. The incubation temperature was at 37°C before the in ocuppation with Myco. lepraemurium, and at 30°C after the inoculation with Myco. lepraemurium. The CO2 content inside the CO2 humidity incubator for the cultivation of macrophage was kept at 5%.
2. In cultures of macrophage inoculated with live Myco. lepraemurium, there observed clear features of incresaes in the number of AF bacilli inside individual cell, elongation of bacilli and increased solidness in AF staining. 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 courses of 6 to 8 weeks after inoculation has provided a partial but substantial evidences of actual multiplication of Myco. lepraemurium in cultured macrophages.
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