Abstract
Mycobacterium leprae, etiologic agent of leprosy, is propagated in athymic nude mouse footpads (FPs). The current purification protocol is tedious and physically demanding. A simpler, semi-automated protocol was developed using gentleMACS™ Octo Dissociator. The gentleMACS protocol provided a very effective means for purification of highly viable M. leprae from tissue.
Highlights
Mycobacterium leprae, etiologic agent of leprosy, is propagated in athymic nude mouse footpads (FPs)
The preferred system for cultivation of highly viable M. leprae for research purposes is the athymic nude mouse footpad (FP) model consisting of injecting M. leprae into each of the hind FP of nude mice and purification of bacteria from FP tissues 5–7 months postinfection (Truman and Krahenbuhl, 2001)
The hand-held homogenization protocol produced ~ 2-fold more M. leprae than the gentleMACS protocol according to acid-fast stain counts (p-value = 0.03) (Truman and Gillis, 2000) (Table 1)
Summary
Mycobacterium leprae, etiologic agent of leprosy, is propagated in athymic nude mouse footpads (FPs). The preferred system for cultivation of highly viable M. leprae for research purposes is the athymic nude mouse footpad (FP) model consisting of injecting M. leprae into each of the hind FP of nude mice and purification of bacteria from FP tissues 5–7 months postinfection (Truman and Krahenbuhl, 2001). The current protocol for purification of highly viable bacteria from this model requires extensive mincing of the tissue with scissors, the production of a tissue homogenate using a hand-held glass tissue grinder and differential centrifugation (Truman and Krahenbuhl, 2001).
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