A rapid rosetting method for separation of hemocyte sub-populations of Drosophila melanogaster

Péter Vilmos, István Nagy, Éva Kurucz, Dan Hultmark, Elisabeth Gateff, István Andó

Research output: Article

25 Citations (Scopus)

Abstract

Hemocytes, cellular elements of the innate immune system in insects, play a crucial role in the cellular and humoral immune response. Although a significant amount of information has been collected on their differentiation and function, our understanding of hemocyte development is far from complete. Their characterisation is mostly based on morphological criteria. However, molecular markers were recently identified, defining functional subsets by the aid of monoclonal antibodies. Isolated subsets of hemocytes, in sufficient quantity and purity could help to analyse their development in vitro and also to further define their molecular characteristics. Here we describe an antibody-based rosetting technique for the physical separation of Drosophila hemocyte sub-populations. We have applied anti-hemocyte antibodies coupled to sheep red blood cells for separation. The method relies on the formation of rosettes between hemocytes and sheep erythrocytes, sensitised with discriminative anti-hemocyte monoclonal antibodies. Using this method the rosetting and non-rosetting hemocytes can be separated from each other by gradient centrifugation. Rosette-forming cells from the pellet and non-rosetting cells from the interface can be isolated in high recovery. The method can be used for functional and molecular characterisation of hemocyte sub-populations. The procedure is sensitive, reproducible and easy to perform.

Original languageEnglish
Pages (from-to)555-563
Number of pages9
JournalDevelopmental and Comparative Immunology
Volume28
Issue number6
DOIs
Publication statusPublished - máj. 17 2004

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ASJC Scopus subject areas

  • Immunology
  • Developmental Biology

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