| Autor: |
Zilberstein D; Faculty of Biology, Technion-Israel Institute of Technology, Haifa, Israel. danz@technion.ac.il., Nitzan Koren R; Faculty of Biology, Technion-Israel Institute of Technology, Haifa, Israel. |
| Jazyk: |
angličtina |
| Zdroj: |
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2019; Vol. 1971, pp. 1-8. |
| DOI: |
10.1007/978-1-4939-9210-2_1 |
| Abstrakt: |
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 CO 2 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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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