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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
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
0736-6205
387
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
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), 387. (doi:10.2144/btn-2021-0020).

Record type: Article

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

e-pub ahead of print date: 16 July 2021
Keywords: Bronchial epithelium, Microfluidic device, ScRNA-seq, Single cell isolation, Virus

Identifiers

Local EPrints ID: 450473
URI: http://eprints.soton.ac.uk/id/eprint/450473
ISSN: 0736-6205
PURE UUID: 676c1788-a754-4929-bbed-c3c4925452fe
ORCID for John Holloway: ORCID iD orcid.org/0000-0001-9998-0464
ORCID for Emily Swindle: ORCID iD orcid.org/0000-0003-3644-7747
ORCID for Matthew Rose-Zerilli: ORCID iD orcid.org/0000-0002-1064-5350

Catalogue record

Date deposited: 29 Jul 2021 16:30
Last modified: 26 Nov 2021 02:56

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Contributors

Author: Lucy M Kimbley
Author: Rachel Parker
Author: Maaike SA Jongen
Author: John Holloway ORCID iD
Author: Emily Swindle ORCID iD

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