Enrichment of Undifferentiated Germline and Somatic Cells from Drosophila Testes
- PMID: 37464239
- DOI: 10.1007/978-1-0716-3259-8_7
Enrichment of Undifferentiated Germline and Somatic Cells from Drosophila Testes
Abstract
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.
Keywords: Drosophila testis; Flow cytometry and cell sorting.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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References
-
- Kaya-Okur HS, Wu SJ, Codomo CA, Pledger ES, Bryson TD, Henikoff JG et al (2019) CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nat Commun 10(1):1930. https://doi.org/10.1038/s41467-019-09982-5 - DOI - PubMed - PMC
-
- Hicks SC, Townes FW, Teng M, Irizarry RA (2018) Missing data and technical variability in single-cell RNA-sequencing experiments. Biostatistics 19(4):562–578. https://doi.org/10.1093/biostatistics/kxx053 - DOI - PubMed
-
- de Cuevas M, Matunis EL (2011) The stem cell niche: lessons from the drosophila testis. Development 138(14):2861–2869. https://doi.org/10.1242/dev.056242 - DOI - PubMed - PMC
-
- Yamashita YM, Jones DL, Fuller MT (2003) Orientation of asymmetric stem cell division by the APC tumor suppressor and centrosome. Science 301(5639):1547–1550. https://doi.org/10.1126/science.1087795 - DOI - PubMed