Methods for cell isolation and analysis of the highly regenerative tunicate Polycarpa mytiligera

Tal Gordon, Noam Hendin, Omri Wurtzel*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Background: Polycarpa mytiligera is the only molecularly characterized solitary ascidian capable of regenerating all organs and tissue types. The cellular basis for regeneration in P. mytiligera is largely unknown, and methods for isolating live cells from this species for functional analyses are unavailable. Results: Here, we developed a method for isolating live cells from P. mytiligera, overcoming major experimental challenges, including the dissociation of its thick body wall and native cellular autofluorescence. We demonstrated the applicability of our approach for tissue dissociation and cell analysis using three flow cytometry platforms, and by using broadly used non-species-specific cell labeling reagents. In addition to live cell isolation, proof-of-concept experiments showed that this approach was compatible with gene expression analysis of RNA extracted from the isolated cells, and with ex vivo analysis of phagocytosis. Conclusion: We presented efficient methods for cell purification from a highly regenerative ascidian, which could be transferable to diversity of non-model marine organisms. The ability to purify live cells will promote future studies of cell function in P. mytiligera regeneration.

Original languageEnglish
Article number1274826
JournalFrontiers in Cell and Developmental Biology
Volume11
DOIs
StatePublished - 2023

Funding

FundersFunder number
Israeli Ministry of Science
European Research Council853640
Israel Science Foundation2039/18
Tel Aviv University

    Keywords

    • autofluorescence
    • cell isolation
    • flow cytometry
    • marine model organisms
    • regeneration

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