Non-invasive harvesting of conjunctival cells for whole transcriptome sequencing

Exp Eye Res. 2023 Sep:234:109613. doi: 10.1016/j.exer.2023.109613. Epub 2023 Aug 12.

Abstract

The purpose of this study was to determine the feasibility of using a non-invasive technique, the EYEPRIM™ conjunctival cell impression device, to harvest sufficient RNA from conjunctival cells for the whole-transcriptome sequencing. Conjunctival cells from 40 participants were collected using an EYEPRIM™ conjunctival cell impression device. RNA was extracted from the samples, followed by library construction and transcriptome sequencing. Quality checks were performed for each technical step of the experiment, and the feasibility of this procedure was examined. RNA of sufficient yield and quality was successfully extracted following additional disruption and homogenization of the conjunctival cells and collection of two impression samples per eye. Successful library preparation and RNA sequencing were performed, with all 40 samples passing the various quality checks used for each step. In conclusion, harvesting cells from the ocular surface using an impression cytology device yields good quality and sufficient mRNA for whole transcriptome sequencing to study diseases of the eye. This technique provides a convenient alternative to using post-mortem tissues or surgical excisions.

Keywords: Conjunctiva; Genetics; Ocular diseases; Transcriptome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Conjunctiva* / metabolism
  • Exome Sequencing
  • Humans
  • RNA* / genetics
  • RNA* / metabolism
  • RNA, Messenger / metabolism

Substances

  • RNA
  • RNA, Messenger