Department of Cell Biology, University of Connecticut Health, Farmington, CT, USA.
Methods Mol Biol. 2023;2677:127-138. doi: 10.1007/978-1-0716-3259-8_7.
The Drosophila male germline provides a strong model system to understand numerous developmental and cell-biological processes, owing to a well-defined anatomy and cell type markers in combination with various genetic tools available for the Drosophila system. A major weakness of this system has been the difficulty of approaches for obtaining material for biochemical assays, proteomics, and genomic or transcriptomic profiling due to small-size and complex tissues. However, the recent development of techniques has started allowing us the usage of a low amount of material for these analyses and now we can strategize many new experiments. The method for enrichment or isolation of rare populations of cells is still challenging and should meaningfully influence the reliability of the results. Here, we provide our semi-optimized protocol of enrichment of undifferentiated germ cells and somatic cells from non-tumorous Drosophila testis, which we have successfully improved after multiple trials.
果蝇雄性生殖细胞为理解众多发育和细胞生物学过程提供了一个强有力的模型系统,这要归功于明确的解剖结构和细胞类型标记,以及可用于果蝇系统的各种遗传工具。由于组织小且复杂,该系统的一个主要弱点一直是获取用于生化分析、蛋白质组学、基因组或转录组分析的材料的方法存在困难。然而,最近技术的发展已经开始允许我们使用少量材料进行这些分析,现在我们可以设计许多新的实验。富集或分离稀有细胞群体的方法仍然具有挑战性,并且应该对结果的可靠性产生有意义的影响。在这里,我们提供了一种从非肿瘤性果蝇睾丸中富集未分化生殖细胞和体细胞的半优化方案,该方案是我们在多次尝试后成功改进的。