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Purification of Planarian Stem Cells Using a Draq5-Based FACS Approach

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Tissue Morphogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2805))

Abstract

Planarians are flatworms that have the remarkable ability to regenerate entirely new animals. This regenerative ability requires abundant adult stem cells called neoblasts, which are relatively small in size, sensitive to irradiation and the only proliferative cells in the animal. Despite the lack of cell surface markers, fluorescence-activated cell sorting (FACS) protocols have been developed to discriminate and isolate neoblasts, based on DNA content. Here, we describe a protocol that combines staining of far-red DNA dye Draq5, Calcein-AM and DAPI, along with a shortened processing time. This profiling strategy can be used to functionally characterize the neoblast population in pharmacologically-treated or gene knockdown animals. Highly purified neoblasts can be analyzed with downstream assays, such as in situ hybridization and RNA sequencing.

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Authors and Affiliations

  1. Department of Molecular Medicine, Cornell University, College of Veterinary Medicine, Ithaca, NY, USA

    Kuang-Tse Wang, Justin Tapper & Carolyn E. Adler

Authors
  1. Kuang-Tse Wang
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  2. Justin Tapper
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  3. Carolyn E. Adler
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Corresponding author

Correspondence to Carolyn E. Adler .

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Editors and Affiliations

  1. 303 Hoyt Laboratory, Princeton University, PRINCETON, NJ, USA

    Celeste M. Nelson

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© 2024 The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Wang, KT., Tapper, J., Adler, C.E. (2024). Purification of Planarian Stem Cells Using a Draq5-Based FACS Approach. In: Nelson, C.M. (eds) Tissue Morphogenesis. Methods in Molecular Biology, vol 2805. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3854-5_14

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  • DOI: https://doi.org/10.1007/978-1-0716-3854-5_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3853-8

  • Online ISBN: 978-1-0716-3854-5

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