Issue 52, 2021

An ultrafast enzyme-free acoustic technique for detaching adhered cells in microchannels

Abstract

Adherent cultured cells are widely used biological tools for a variety of biochemical and biotechnology applications, including drug screening and gene expression analysis. One critical step in culturing adherent cells is the dissociation of cell monolayers into single-cell suspensions. Different enzymatic and non-enzymatic methods have been proposed for this purpose. Trypsinization, the most common enzymatic method for dislodging adhered cells, can be detrimental to cells, as it can damage cell membranes and ultimately cause cell death. Additionally, all available techniques require a prolonged treatment duration, typically on the order of minutes (5–10 min). Dissociation of cells becomes even more challenging in microfluidic devices, where, due to the nature of low Reynolds number flow and reduced mixing efficiency, multiple washing steps and prolonged trypsinization may be necessary to treat all cells. Here, we report a novel acoustofluidic method for the detachment of cells adhered onto a microchannel surface without exposing the cells to any enzymatic or non-enzymatic chemicals. This method enables a rapid (i.e., on the order of seconds), cost-effective, and easy-to-operate cell detachment strategy, yielding a detachment efficiency of ∼99% and cellular viability similar to that of the conventional trypsinization method. Also, as opposed to biochemical-based techniques (e.g., enzymatic), in our approach, cells are exposed to the dissociating agent (i.e., substrate-mediated acoustic excitation and microstreaming flow) only for as long as they remain attached to the substrate. After dissociation, the effect of acoustic excitation is reduced to microstreaming flow, therefore, minimizing unwanted effects of the dissociating agent on the cell phenotype. Additionally, our results suggest that cell excitation at acoustic powers lower than that required for complete cell detachment can potentially be employed for probing the adhesion strength of cell–substrate attachment. This novel approach can, therefore, be used for a wide range of lab-on-a-chip applications.

Graphical abstract: An ultrafast enzyme-free acoustic technique for detaching adhered cells in microchannels

Supplementary files

Article information

Article type
Paper
Submitted
23 jun 2021
Accepted
26 sep 2021
First published
05 okt 2021
This article is Open Access
Creative Commons BY-NC license

RSC Adv., 2021,11, 32824-32829

An ultrafast enzyme-free acoustic technique for detaching adhered cells in microchannels

A. Salari, S. Appak-Baskoy, I. R. Coe, S. S. H. Tsai and M. C. Kolios, RSC Adv., 2021, 11, 32824 DOI: 10.1039/D1RA04875A

This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. You can use material from this article in other publications, without requesting further permission from the RSC, provided that the correct acknowledgement is given and it is not used for commercial purposes.

To request permission to reproduce material from this article in a commercial publication, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party commercial publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements