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A Microfluidic Platform for High-throughput Single-cell Isolation and Culture

A Microfluidic Platform for High-throughput Single-cell Isolation and Culture

Published on: June 16, 2016

Selective single cell isolation for genomics using microraft arrays.

Joshua D Welch¹, Lindsay A Williams², Matthew DiSalvo³

¹Curriculum in Bioinformatics and Computational Biology, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-3280, USA Department of Computer Science, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-3280, USA.

Nucleic Acids Research

|August 18, 2016

Summary

This summary is machine-generated.

Microraft arrays enable researchers to isolate single cells based on diverse phenotypes for genomic analysis. This technique, coupled with RNA-sequencing, successfully identified gene expression changes linked to early drug resistance in pancreatic cancer cells.

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Area of Science:

Single-cell genomics
Cellular phenotyping
Cancer biology

Background:

Genomic methods are increasingly used to study individual cells within tissues.
Current single-cell isolation methods often rely on fluorescent markers and struggle with complex phenotypes.
Connecting genotype to phenotype at the single-cell level remains challenging.

Purpose of the Study:

To develop a method for phenotypically selecting single cells for genomic analysis.
To adapt RNA-sequencing for use with single cells isolated from microraft arrays.
To investigate gene expression changes associated with drug resistance in pancreatic cancer.

Main Methods:

Utilized microraft arrays for high-throughput single-cell culture and phenotypic selection.
Adapted RNA-sequencing (RNA-seq) for analyzing RNA from single cells isolated on microrafts.
Compared microraft-based RNA-seq with the Fluidigm C1 platform.

Main Results:

Microraft arrays successfully selected single cells based on various phenotypes, including drug response.
The adapted RNA-seq protocol yielded high-quality data comparable to existing methods.
Identified novel and known gene expression alterations in drug-resistant pancreatic cancer cells.

Conclusions:

Microraft arrays provide a versatile platform for phenotypically driven single-cell genomic studies.
This approach facilitates the connection between genotype and complex cellular phenotypes.
The findings offer insights into the molecular mechanisms of early drug resistance in pancreatic cancer.