Continuous muscle, glial, epithelial, neuronal, and hemocyte cell lines for Drosophila research

  1. Nikki Coleman-Gosser
  2. Yanhui Hu
  3. Shiva Raghuvanshi
  4. Shane Stitzinger
  5. Weihang Chen
  6. Arthur Luhur
  7. Daniel Mariyappa
  8. Molly Josifov
  9. Andrew Zelhof
  10. Stephanie E Mohr
  11. Norbert Perrimon  Is a corresponding author
  12. Amanda Simcox  Is a corresponding author
  1. The Ohio State University, United States
  2. Harvard Medical School, United States
  3. Indiana University, United States
  4. Harvard Medical School, Howard Hughes Medical Institute, United States

Abstract

Expression of activated Ras, RasV12, provides Drosophila cultured cells with a proliferation and survival advantage that simplifies the generation of continuous cell lines. Here we used lineage restricted RasV12 expression to generate continuous cell lines of muscle, glial, and epithelial cell type. Additionally, cell lines with neuronal and hemocyte characteristics were isolated by cloning from cell cultures established with broad RasV12 expression. Differentiation with the hormone ecdysone caused maturation of cells from mesoderm lines into active muscle tissue and enhanced dendritic features in neuronal-like lines. Transcriptome analysis showed expression of key cell-type specific genes and the expected alignment with single cell sequencing and in situ data. Overall, the technique has produced in vitro cell models with characteristics of glia, epithelium, muscle, nerve, and hemocyte. The cells and associated data are available from the Drosophila Genomic Resource Center.

Data availability

Sequencing data have been deposited in GEO under accession code GSE219105.

The following data sets were generated

Article and author information

Author details

  1. Nikki Coleman-Gosser

    Department of Molecular Genetics, The Ohio State University, Columbus, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Yanhui Hu

    Department of Genetics, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Shiva Raghuvanshi

    Department of Molecular Genetics, The Ohio State University, Columbus, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Shane Stitzinger

    Department of Molecular Genetics, The Ohio State University, Columbus, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Weihang Chen

    Department of Genetics, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Arthur Luhur

    Department of Biology, Indiana University, Bloomington, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Daniel Mariyappa

    Department of Biology, Indiana University, Bloomington, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4775-1656
  8. Molly Josifov

    Department of Molecular Genetics, The Ohio State University, Columbus, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2899-7186
  9. Andrew Zelhof

    Department of Biology, Indiana University, Bloomington, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Stephanie E Mohr

    Department of Genetics, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9639-7708
  11. Norbert Perrimon

    Harvard Medical School, Howard Hughes Medical Institute, Boston, United States
    For correspondence
    perrimon@genetics.med.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7542-472X
  12. Amanda Simcox

    Department of Molecular Genetics, The Ohio State University, Columbus, United States
    For correspondence
    simcox.1@osu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5572-7042

Funding

National Institutes of Health Office of the Director (R24 OD019847)

  • Stephanie E Mohr
  • Norbert Perrimon
  • Amanda Simcox

National Institutes of Health (P40OD010949)

  • Andrew Zelhof

National Institutes of Health (P41 GM132087)

  • Andrew Zelhof

National Science Foundation (IOS 1419535)

  • Amanda Simcox

Howard Hughes Medical Institute

  • Norbert Perrimon

Women & Philanthropy at the Ohio State University (Grant)

  • Amanda Simcox

National Science Foundation (Support while serving at the National Science Foundation)

  • Amanda Simcox

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Any opinion, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Science Foundation.

Reviewing Editor

  1. Erika A Bach, New York University School of Medicine, United States

Version history

  1. Received: December 28, 2022
  2. Preprint posted: January 19, 2023 (view preprint)
  3. Accepted: July 12, 2023
  4. Accepted Manuscript published: July 20, 2023 (version 1)
  5. Version of Record published: August 1, 2023 (version 2)

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Metrics

  • 1,624
    views
  • 318
    downloads
  • 0
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Nikki Coleman-Gosser
  2. Yanhui Hu
  3. Shiva Raghuvanshi
  4. Shane Stitzinger
  5. Weihang Chen
  6. Arthur Luhur
  7. Daniel Mariyappa
  8. Molly Josifov
  9. Andrew Zelhof
  10. Stephanie E Mohr
  11. Norbert Perrimon
  12. Amanda Simcox
(2023)
Continuous muscle, glial, epithelial, neuronal, and hemocyte cell lines for Drosophila research
eLife 12:e85814.
https://doi.org/10.7554/eLife.85814

Share this article

https://doi.org/10.7554/eLife.85814

Further reading

    1. Cell Biology
    Yoko Nakai-Futatsugi, Jianshi Jin ... Masayo Takahashi
    Research Article

    Retinal pigment epithelium (RPE) cells show heterogeneous levels of pigmentation when cultured in vitro. To know whether their color in appearance is correlated with the function of the RPE, we analyzed the color intensities of human-induced pluripotent stem cell-derived RPE cells (iPSC-RPE) together with the gene expression profile at the single-cell level. For this purpose, we utilized our recent invention, Automated Live imaging and cell Picking System (ALPS), which enabled photographing each cell before RNA-sequencing analysis to profile the gene expression of each cell. While our iPSC-RPE were categorized into four clusters by gene expression, the color intensity of iPSC-RPE did not project any specific gene expression profiles. We reasoned this by less correlation between the actual color and the gene expressions that directly define the level of pigmentation, from which we hypothesized the color of RPE cells may be a temporal condition not strongly indicating the functional characteristics of the RPE.

    1. Cancer Biology
    2. Cell Biology
    Savvas Nikolaou, Amelie Juin ... Laura M Machesky
    Research Article

    Pancreatic ductal adenocarcinoma carries a dismal prognosis, with high rates of metastasis and few treatment options. Hyperactivation of KRAS in almost all tumours drives RAC1 activation, conferring enhanced migratory and proliferative capacity as well as macropinocytosis. Macropinocytosis is well understood as a nutrient scavenging mechanism, but little is known about its functions in trafficking of signaling receptors. We find that CYRI-B is highly expressed in pancreatic tumours in a mouse model of KRAS and p53-driven pancreatic cancer. Deletion of Cyrib (the gene encoding CYRI-B protein) accelerates tumourigenesis, leading to enhanced ERK and JNK-induced proliferation in precancerous lesions, indicating a potential role as a buffer of RAC1 hyperactivation in early stages. However, as disease progresses, loss of CYRI-B inhibits metastasis. CYRI-B depleted tumour cells show reduced chemotactic responses to lysophosphatidic acid, a major driver of tumour spread, due to impaired macropinocytic uptake of the lysophosphatidic acid receptor-1. Overall, we implicate CYRI-B as a mediator of growth and signaling in pancreatic cancer, providing new insights into pathways controlling metastasis.