BioTechniques (Jul 2021)

Rapid microfluidic isolation of virally infected primary bronchial epithelial cells for single-cell RNA sequencing

  • Lucy M Kimbley,
  • Rachel Parker,
  • Maaike Sybil Jongen,
  • John W Holloway,
  • Emily J Swindle,
  • Matthew JJ Rose-Zerilli

DOI
https://doi.org/10.2144/btn-2021-0020
Journal volume & issue
Vol. 71, no. 1
pp. 387 – 391

Abstract

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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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