Papain-Based Dissociation of Schmidtea mediterranea Cells
Autor(en): | Ortmeier, Claudia Gentile, Luca |
Stichwörter: | Cell dissociation; cell selection; cell transplantation; cell viability; cysteine; cysteine proteinase; enzyme metabolism; flow cytometry; gene expression profiling; High cell viability; immunohistochemistry; Mediterranea; nonhuman; Papain; Schmidtea mediterranea; Schmidteamediterranea; transcriptomics | Erscheinungsdatum: | 2023 | Herausgeber: | Humana Press Inc. | Journal: | Methods in Molecular Biology | Volumen: | 2680 | Startseite: | 179 – 187 | Zusammenfassung: | The need of highly viable cellsdissociated from Schmidtea mediterranea is constantly growing. In this chapter, we describe a cell dissociation method based on papain (papaya peptidase I). This enzyme, often used to dissociate cells with complex morphology, is a cysteine protease with a broad specificity and increases both the yield and the viability of the dissociated cell suspension. The papain dissociation is preceded by a pretreatment for mucus removal, as this was shown to greatly improve the yield of cell dissociation, regardless of the method used. Papain-dissociated cells are suitable for a variety of downstream applications, like live immunostaining, flow cytometry, cell sorting, transcriptomics, and cell transplantation, also at the single-cell level. © 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature. |
Beschreibung: | Cited by: 0 |
ISSN: | 1064-3745 | DOI: | 10.1007/978-1-0716-3275-8_11 | Externe URL: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85164286298&doi=10.1007%2f978-1-0716-3275-8_11&partnerID=40&md5=3f0ac70ebd1c3a85ca967f61a4c1b90f |
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