A minimally invasive method for retrieving single adherent cells of different types from cultures

Jia Zeng, Aida Mohammadreza, Weimin Gao, Saeed Merza, Dean Smith, Laimonas Kelbauskas, Deirdre Meldrum

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Abstract

The field of single-cell analysis has gained a significant momentum over the last decade. Separation and isolation of individual cells is an indispensable step in almost all currently available single-cell analysis technologies. However, stress levels introduced by such manipulations remain largely unstudied. We present a method for minimally invasive retrieval of selected individual adherent cells of different types from cell cultures. The method is based on a combination of mechanical (shear flow) force and biochemical (trypsin digestion) treatment. We quantified alterations in the transcription levels of stress response genes in individual cells exposed to varying levels of shear flow and trypsinization. We report optimal temperature, RNA preservation reagents, shear force and trypsinization conditions necessary to minimize changes in the stress-related gene expression levels. The method and experimental findings are broadly applicable and can be used by a broad research community working in the field of single cell analysis.

Original languageEnglish (US)
Article number5424
JournalScientific reports
Volume4
DOIs
StatePublished - Jun 24 2014

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