Rapid microfluidic isolation of virally infected primary bronchial epithelial cells for single-cell RNA sequencing
Rapid microfluidic isolation of virally infected primary bronchial epithelial cells for single-cell RNA sequencing
Single-cell RNA sequencing (scRNA-seq) of the bronchial epithelium enables examination of cellular subtypes and their responses to viral infections. Here, an optimized method for the isolation of virally infected primary bronchial epithelial cells using a commercially available microfluidic device is presented. Using this method single cells can be rapidly isolated with minimal equipment available in most laboratories. Isolation can be carried out inside biological safety cabinets, permitting the use of virally infected cells. Both cell-line and primary cells isolated using the device retained sufficient RNA integrity for the generation of short-read sequencing-compatible cDNA libraries to facilitate scRNA-seq.
Bronchial epithelium, Microfluidic device, ScRNA-seq, Single cell isolation, Virus
387-391
Kimbley, Lucy M
3e287c79-12da-4699-b4b4-d23bd3abd131
Parker, Rachel
ca68b9e6-b3d4-4246-845f-4636068d479f
Jongen, Maaike SA
13f00bd0-5bf2-400f-a3ac-aa90d2180327
Holloway, John
4bbd77e6-c095-445d-a36b-a50a72f6fe1a
Swindle, Emily
fe393c7a-a513-4de4-b02e-27369bd7e84f
Rose-Zerilli, Matthew
08b3afa4-dbc2-4c0d-a852-2a9f33431199
17 July 2021
Kimbley, Lucy M
3e287c79-12da-4699-b4b4-d23bd3abd131
Parker, Rachel
ca68b9e6-b3d4-4246-845f-4636068d479f
Jongen, Maaike SA
13f00bd0-5bf2-400f-a3ac-aa90d2180327
Holloway, John
4bbd77e6-c095-445d-a36b-a50a72f6fe1a
Swindle, Emily
fe393c7a-a513-4de4-b02e-27369bd7e84f
Rose-Zerilli, Matthew
08b3afa4-dbc2-4c0d-a852-2a9f33431199
Kimbley, Lucy M, Parker, Rachel, Jongen, Maaike SA, Holloway, John, Swindle, Emily and Rose-Zerilli, Matthew
(2021)
Rapid microfluidic isolation of virally infected primary bronchial epithelial cells for single-cell RNA sequencing.
Biotechniques, 71 (1), .
(doi:10.2144/btn-2021-0020).
Abstract
Single-cell RNA sequencing (scRNA-seq) of the bronchial epithelium enables examination of cellular subtypes and their responses to viral infections. Here, an optimized method for the isolation of virally infected primary bronchial epithelial cells using a commercially available microfluidic device is presented. Using this method single cells can be rapidly isolated with minimal equipment available in most laboratories. Isolation can be carried out inside biological safety cabinets, permitting the use of virally infected cells. Both cell-line and primary cells isolated using the device retained sufficient RNA integrity for the generation of short-read sequencing-compatible cDNA libraries to facilitate scRNA-seq.
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More information
Accepted/In Press date: 16 June 2021
e-pub ahead of print date: 16 July 2021
Published date: 17 July 2021
Additional Information:
Funding Information:
This work was supported by an Asthma Allergy and Inflammation (AAIR) Charity Research grant and a grant from the Southampton CR UK Development Fund. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.
Publisher Copyright:
© 2021 Matthew Rose-Zerilli.
Keywords:
Bronchial epithelium, Microfluidic device, ScRNA-seq, Single cell isolation, Virus
Identifiers
Local EPrints ID: 456889
URI: http://eprints.soton.ac.uk/id/eprint/456889
ISSN: 0736-6205
PURE UUID: 36d090bb-e791-494c-a068-16c700b56f67
Catalogue record
Date deposited: 16 May 2022 16:31
Last modified: 06 Jun 2024 01:48
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Contributors
Author:
Lucy M Kimbley
Author:
Rachel Parker
Author:
Maaike SA Jongen
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