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

Lucy M KimbleyRachel ParkerMaaike Sybil JongenJohn W HollowayEmily Jane SwindleMatthew J J Rose-Zerilli
Published in: BioTechniques (2021)
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.
Keyphrases
  • single cell
  • rna seq
  • induced apoptosis
  • high throughput
  • cell cycle arrest
  • sars cov
  • cell death
  • circulating tumor cells
  • gene expression
  • dna methylation
  • quantum dots