Generating and working with Drosophila cell cultures: Current challenges and opportunities

Abstract

The use of Drosophila cell cultures has positively impacted both fundamental and biomedical research. The most widely used cell lines: Schneider, Kc, the CNS and imaginal disc lines continue to be the choice for many applications. Drosophila cell lines provide a homogenous source of cells suitable for biochemical experimentations, transcriptomics, functional genomics, and biomedical applications. They are amenable to RNA interference and serve as a platform for high-throughput screens to identify relevant candidate genes or drugs for any biological process. Currently, CRISPR-based functional genomics are also being developed for Drosophila cell lines. Even though many uniquely derived cell lines exist, cell genetic techniques such the transgenic UAS-GAL4-based Ras^V12 oncogene expression, CRISPR-Cas9 editing and recombination mediated cassette exchange are likely to drive the establishment of many more lines from specific tissues, cells, or genotypes. However, the pace of creating new lines is hindered by several factors inherent to working with Drosophila cell cultures: single cell cloning, optimal media formulations and culture conditions capable of supporting lines from novel tissue sources or genotypes. Moreover, even though many Drosophila cell lines are morphologically and transcriptionally distinct it may be necessary to implement a standard for Drosophila cell line authentication, ensuring the identity and purity of each cell line. Altogether, recent advances and a standardized authentication effort should improve the utility of Drosophila cell cultures as a relevant model for fundamental and biomedical research. This article is categorized under: Technologies > Analysis of Cell, Tissue, and Animal Phenotypes.

Keywords: CRISPR; Drosophila; RNA interference; authentication; cell culture.

Figure 1.

Multiple Drosophila cell lines with different morphologies. Round cells of S2-DRSC (A), mbn2 (B), Kc167 (C) and Jupiter-GFP, Dsas-9 cells (H). CNS-derived cell lines ML-BG3-c2 (D) and ML-BG2-c2 (I) show very distinct morphologies. RasV12; wtsRNAi (WRR1) cell line with epithelial appearance (E) and ovaryderived OSS (F) cell line. Larval wing disc Cl.8+ cells (G) with fibroblast-like morphology. Scale bar = 25m (Images courtesy of DGRC, Johnny Roberts).

Figure 2.

(Top) Ideal arrangement of the workspace in the laminar hood and handling one cell line at a time. (Bottom) A cluttered and overcrowded work area in the laminar hood compromises the airflow and thus the sterility in the laminar hood.

Figure 3.

The accession process and cryopreservation of a Drosophila cell line.