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Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening

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Published on: September 18, 2016

Single-cell analysis of embryoid body heterogeneity using microfluidic trapping array.

Jenna L Wilson¹, Shalu Suri², Ankur Singh³

¹The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.

Biomedical Microdevices

|October 3, 2013

Summary

This summary is machine-generated.

This study introduces a microfluidic cell trapping array to analyze cell heterogeneity in embryoid bodies (EBs) during differentiation. The novel method accurately assesses single-cell phenotypes, revealing differentiation patterns and pluripotency loss.

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Area of Science:

Stem cell biology
Developmental biology
Biotechnology

Background:

Embryoid bodies (EBs) are widely used for pluripotent stem cell differentiation but often result in heterogeneous cell populations.
Physical factors like EB size influence differentiation heterogeneity due to nutrient and oxygen gradients.
Analyzing single-cell phenotypes within EBs is challenging due to a lack of suitable analytical methods.

Purpose of the Study:

To evaluate a microfluidic cell trapping array for analyzing cellular heterogeneity in EBs during early differentiation.
To assess single-cell phenotypes based on OCT-4 protein expression in EBs of varying sizes and differentiation stages.

Main Methods:

Utilized a microfluidic cell trapping array to analyze individual cells from EBs.
Examined OCT-4 protein expression as a marker for pluripotency and differentiation.
Compared results with traditional methods like flow cytometry and whole mount immunostaining.

Main Results:

The microfluidic array successfully analyzed single-cell heterogeneity in EBs.
Demonstrated the ability to discern differences in OCT-4 expression between pooled and single EB analyses.
Provided insights into how EB size and differentiation stage impact cellular heterogeneity.

Conclusions:

The microfluidic cell trapping array offers a novel approach to evaluate EB differentiation heterogeneity.
This method can assess differentiation kinetics, patterns, and pluripotency loss at the single-cell level.
Future applications include optimizing differentiation protocols and understanding developmental processes.