Host-Free Systems for Differentiation of Axenic Leishmania

  • Dan ZilbersteinEmail author
  • Roni Nitzan Koren
Part of the Methods in Molecular Biology book series (MIMB, volume 1971)


This chapter describes, in detail, the method our laboratory developed to differentiate L. donovani promastigotes into amastigotes in a host-free culture. This method is based on previous observations that Leishmania promastigotes can combine two environmental signals, typical to lysosomes, acidic pH (~5.5) and body temperature (37 °C), into a signal that induces differentiation. Based on this concept, we have modified medium 199 to make it into an amastigote-specific medium. Shifting promastigotes to this medium, followed by incubation in a CO2 incubator, induced differentiation. Axenic amastigotes reach maturation within 5 days, resembling the time it takes in vivo. This chapter provides a complete protocol that should be useful for both Old and New World species of Leishmania.

Key words

Development Phagolysosome Differentiation Signaling Promastigote Amastigote 


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Faculty of BiologyTechnion-Israel Institute of TechnologyHaifaIsrael

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