Ultrastructural Characterization of Serially Passaged Amastigote Like Forms of Leishmania (Leishmania) Donovani

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The present study was done to establish an in vitro axenic culture of amastigote like forms of Leishmania (Leishmania) donovani (Dd-8 strain), the causative agent of Indian kala-azar. Transformation of promastigotes to amastigote like forms was induced by temperature shift from 26±1℃ to 34±1℃ at pH 7.0 in NNN medium. These forms were dividing as evidenced by flow cytometry. Scanning and transmission electron microscopic studies revealed a remarkable ultrastructural similarity of these in vitro cultured amastigotes with intracellular amastigotes. These forms have been successfully maintained for a period of more than one year, during which they have remained infective. On subjecting these forms to temperature of 26±1℃, they reverted back to the promastigote forms. Thus a simple NNN medium, free from foetal calf serum has been developed to generate large amounts of amastigote like forms which can be used for further biochemical, immunological and chemotherapeutic studies.
長崎大学の論文