Generating and working with Drosophila cell cultures: Current challenges and opportunities
Generating and working with Drosophila cell cultures: Current challenges and opportunities
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DOI：
10.1002/wdev.339
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发表时间：
2019-05-01
影响因子：
--
通讯作者：
Zelhof, Andrew C.
中科院分区：
文献类型：
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作者：
Luhur, Arthur;Klueg, Kristin M.;Zelhof, Andrew C.
果蝇细胞培养物的使用对基础研究和生物医学研究都产生了积极的影响。最广泛使用的细胞系：Schneider、Kc、CNS和成虫盘细胞系仍然是许多应用的选择。果蝇细胞系为生物化学实验、转录组学、功能基因组学和生物医学应用提供了均一的细胞来源。它们适合RNA干扰，并作为高通量筛选的平台，以识别任何生物过程的相关候选基因或药物。目前，基于CRISPR的功能基因组学也正在为果蝇细胞系开发。尽管存在许多独特衍生的细胞系，但细胞遗传技术，如基于UAS-GAL 4的转基因Ras（V12）癌基因表达、CRISPR-Cas9编辑和重组介导的盒交换，可能会推动从特定组织、细胞或基因型建立更多的细胞系。然而，创建新品系的步伐受到果蝇细胞培养固有的几个因素的阻碍：单细胞克隆，最佳培养基配方和培养条件能够支持来自新组织来源或基因型的品系。此外，尽管许多果蝇细胞系在形态和转录上是不同的，但可能有必要实施果蝇细胞系认证的标准，以确保每个细胞系的身份和纯度。总之，最新的进展和标准化的认证工作应提高果蝇细胞培养物作为基础和生物医学研究的相关模型的实用性。
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.