Protocol for culturing and functionally manipulating planarian neoblasts using SiR-DNA-based flow cytometry

Bibliographic Details
Title: Protocol for culturing and functionally manipulating planarian neoblasts using SiR-DNA-based flow cytometry
Authors: Wenya Zhang, Xinran Li, Yun Zhao, Kai Lei
Source: STAR Protocols, Vol 5, Iss 1, Pp 102877- (2024)
Publisher Information: Elsevier, 2024.
Publication Year: 2024
Collection: LCC:Science (General)
Subject Terms: Cell culture, Cell isolation, Developmental biology, Model Organisms, Stem Cells, Science (General), Q1-390
More Details: Summary: Neoblasts are the only cells capable of proliferation in planarians. The traditional flow cytometry protocol using Hoechst inhibits the cell cycle. Here, we present a protocol for culturing and functionally manipulating planarian neoblasts using SiR-DNA-based flow cytometry. We describe steps for cell dissociation and staining, flow cytometry, and cell collection and culture. We then detail procedures for Nanoluciferase mRNA transfection. This protocol facilitates further investigations into the pluripotency and regeneration mechanisms within neoblasts.For complete details on the use and execution of this protocol, please refer to Lei et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Document Type: article
File Description: electronic resource
Language: English
ISSN: 2666-1667
Relation: http://www.sciencedirect.com/science/article/pii/S266616672400042X; https://doaj.org/toc/2666-1667
DOI: 10.1016/j.xpro.2024.102877
Access URL: https://doaj.org/article/df356731ef9c4fd4902451f7dda9bb64
Accession Number: edsdoj.f356731ef9c4fd4902451f7dda9bb64
Database: Directory of Open Access Journals
More Details
ISSN:26661667
DOI:10.1016/j.xpro.2024.102877
Published in:STAR Protocols
Language:English