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KMID : 0377619680140040293
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Korean Jungang Medical Journal
1968 Volume.14 No. 4 p.293 ~ p.297
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An Attempt to Cultivate Mycobacterium leprae in Cultured Rabbit Embryo Fibroblast (REF)
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梁容泰/Yang, Y.T.
柳駿/張在薰/Lew, J./Chong, C.H.
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Abstract
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Multiplication of Mycobacterium leprae was attempted by inoculation of human leprosy bacilli into the cultured rabbit embryo fibroblast (REF) that had been originated and subcultured by serial passages at the Department since July 1965. REF was cultured on the cover slip inside the Leighton tube, and suspensions of human leprosy bacilli were prepared from the biopsied nodules from fresh lepromatous leprosy patients who made registration at the World Vision Leprosy Research Center. The inoculated cultures were incubated at 37°C under 5% CO2 atomosphere for the maximum period of 60 days after inoculation. Microscopic observation of the slide prepared and stained for acid-fast bacilli at appropriate intervals after inoculation was followed, and the results are summarized and concluded as follows:
1. REF ingested human leprosy bacilli well. However, uptake of human leprosy bacilli by cultured REF appeared to be a very slow process, and even after the contact period of 48 hours the overlaying medium did contain a large number of the bacilli. Addition of bovine or horse serum to the tissue culture medium exerted a similar effectiveness in the process Of uptake of leprosy bacilli by REF.
2. During the observation period of up to 60 days after the inoculation of leprosy bacilli into the cultured REF, there observed that individual REF contained significantly increased number of acid-fast staining bacilli. It was also possible to observe features of elongation of individual acid-fast staining bacilli inside the cells.
3. Because of relatively short period of observation after inoculation with leprosy bacilli, and of occurrence of nonspecific spontaneous deterioration of REF which developed through the prolonged time of incubation, it was impossible to assess the actual multiplication of human leprosy bacilli in cultured REF.
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