Authors
Zachary M Lane, Christine E Schnitzler
Published in
bioRxiv : the preprint server for biology. Jul 23, 2026. Epub Jul 23, 2026.
Abstract
Hydractinia symbiolongicarpus is a powerful model for stem cell research and maintains a population of pluripotent adult stem cells throughout its lifetime. Here we describe a gene expression-agnostic FACS technique to isolate a live cell population from Hydractinia feeding polyps that appear to be stem cells. This technique utilizes only the general cellular component stains DAPI, DRAQ5, Calcein AM, and Pyronin Y. The stem cell population was identified via subtractive gating based on samples whose stem cell populations had been selectively depleted with the DNA-alkylating agent Mitomycin C. To validate the identity of the isolated population, a colorimetric cytological assay capable of simultaneously discriminating between all major Hydractinia cell types in a live-dissociated cell solution was developed using May-Grünwald and Giemsa stains. The isolated cell population was significantly depleted by Mitomycin C administration, had a high RNA content, was proliferative, had a cytological profile that matched that of Piwi1 + stem cells, and was ∼10x enriched with Piwi1 + stem cells compared to whole cell suspension, all of which support the conclusion that the isolated population is indeed comprised of stem cells. This gene-agnostic FACS technique will serve future research into Hydractinia stem cell biology by enabling the use of isolated populations of live stem cells in transplantation, cell culture, and spheroid experimentation, and may serve as a reference for the development of new methods in other cnidarian species.
PMID:
42539168
Bibliographic data and abstract were imported from PubMed on 01 Aug 2026.
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