Preparing a Single Cell Suspension from Zebrafish Retinal Tissue for Flow Cytometric Cell Sorting of Müller Glia

Preparation of a single cell suspension from solid tissue is vital for a successful flow cytometry experiment. We report a detailed and reproducible method to produce a quality cell suspension from the zebrafish retina. Zebrafish retinas, especially their Müller glia cells, are of particular interes...

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Bibliographic Details
Published in:Cytometry. Part A 2020-06, Vol.97 (6), p.638-646
Main Authors: Allan, Kristin, DiCicco, Rose, Ramos, Michael, Asosingh, Kewal, Yuan, Alex
Format: Article
Language:English
Subjects:
Apolipoprotein E
Cell suspensions
Cell therapy
Danio rerio
dissociation
flow cytometric cell sorting
Flow cytometry
Müller glia
Papain
Regenerative medicine
Retina
Tissue engineering
Tissues
Zebrafish
zebrafish retina
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Summary:Preparation of a single cell suspension from solid tissue is vital for a successful flow cytometry experiment. We report a detailed and reproducible method to produce a quality cell suspension from the zebrafish retina. Zebrafish retinas, especially their Müller glia cells, are of particular interest for their inherent regenerative capacity, making them a useful model for regenerative medicine and cell therapy research. Here, we detail a papain‐based dissociation that is gentle enough to keep cells intact, but strong enough to disrupt cell‐cell and cell‐matrix interactions to yield a cell suspension that produces clean and reliable flow cytometric cell sorting results. This procedure consistently results in over 90% viability and three populations of cells based on GFP expression. The dissociation procedure described herein has been optimized for the collection of Müller glia from Tg(apoe:gfp) zebrafish retinas; however, the overall process may be applicable to other cell types in the fish retina, additional flow cytometric techniques, or preparing cell suspensions from similar tissues. © 2019 International Society for Advancement of Cytometry
ISSN:1552-4922
1552-4930
DOI:10.1002/cyto.a.23936