Dhople A M, Green K J, Osborne L J
Medical Research Institute, Florida Institute of Technology, Melbourne 32904.
Ann Inst Pasteur Microbiol. 1988 Mar-Apr;139(2):213-23. doi: 10.1016/0769-2609(88)90006-3.
The inability to cultivate Mycobacterium leprae in vitro has been a major bottleneck in leprosy research. There have been numerous reports on successful in vitro cultivation of this organism, but these reports could not be confirmed by others in the field. Hence, in vitro multiplication of M. leprae was evaluated in various culture media. Only 2 media supported limited multiplication of M. leprae. One medium was used previously by one of the authors (AMD) for in vitro growth of M. lepraemurium and the other was a conditioned medium used for growth of mouse dorsal root ganglion. Growth was evaluated by 3 biochemical parameters: bacterial ATP, DNA and 3H-thymidine uptake. All 3 measurements revealed a 4-6-fold increase in cell biomass after 16 weeks of incubation at 34 degrees C. The harvested bacilli demonstrated a few of the important properties of M. leprae, including growth in mouse footpads. However, subcultures of these in-vitro-grown cells in the respective media could not be achieved. By the end of 12 weeks, the bacilli lost all intracellular ATP and the ability to incorporate 3H-thymidine; they also failed to multiply in mouse footpads.
无法在体外培养麻风分枝杆菌一直是麻风病研究中的一个主要瓶颈。关于该生物体成功进行体外培养已有众多报道，但这些报道无法得到该领域其他研究者的证实。因此，在各种培养基中对麻风分枝杆菌的体外增殖进行了评估。只有两种培养基支持麻风分枝杆菌的有限增殖。其中一种培养基曾被作者之一（AMD）用于鼠麻风分枝杆菌的体外生长，另一种是用于小鼠背根神经节生长的条件培养基。通过三个生化参数评估生长情况：细菌ATP、DNA以及3H-胸腺嘧啶核苷摄取。所有这三项测量均显示，在34摄氏度孵育16周后，细胞生物量增加了4至6倍。收获的杆菌表现出了麻风分枝杆菌的一些重要特性，包括在小鼠足垫中生长。然而，无法在各自的培养基中对这些体外生长的细胞进行传代培养。到12周结束时，杆菌失去了所有细胞内ATP以及掺入3H-胸腺嘧啶核苷的能力；它们也无法在小鼠足垫中增殖。
