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A red-shifted fluorescent substrate for aldehyde dehydrogenase.

Il Minn¹, Haofan Wang¹, Ronnie C Mease²

¹1] Russel H. Morgan Department of Radiology and Radiological Science, Baltimore, Maryland 21287, USA [2].

Nature Communications

|April 25, 2014

Summary

A new red-shifted fluorescent substrate, AldeRed 588-A, enables researchers to easily identify and isolate aldehyde dehydrogenase (ALDH)-positive cells. This tool is effective for stem cell selection and studying ALDH activity, even with green fluorescent markers.

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Area of Science:

Biochemistry
Cell Biology
Stem Cell Research

Background:

Selecting aldehyde dehydrogenase (ALDH)-positive cells is challenging due to interference from green fluorescent markers.
Existing reagents often struggle to differentiate ALDH-positive cells in complex biological samples.

Purpose of the Study:

To develop and validate a novel red-shifted fluorescent substrate for aldehyde dehydrogenase (ALDH) activity.
To enable the efficient isolation and study of viable ALDH-positive cells, particularly in the presence of green fluorophores.

Main Methods:

Development of AldeRed 588-A, a red-shifted fluorescent substrate for ALDH.
Application of AldeRed 588-A for isolating ALDH-high (ALDHhi) human hematopoietic stem cells from cord blood.
Utilizing AldeRed 588-A in multicolour applications with green fluorophores like ALDEFLUOR and enhanced green-fluorescent protein (eGFP).
Microscopic visualization of ALDH-positive murine pancreatic cells stained with AldeRed 588-A.

Main Results:

AldeRed 588-A successfully labels viable ALDH-positive cells.
Efficient isolation of ALDH-high human hematopoietic stem cells from heterogeneous cord blood mononuclear cells was achieved.
AldeRed 588-A demonstrated compatibility with multicolour applications, allowing ALDH-positive cell fractionation alongside green fluorescent markers.
Successful staining of ALDH-positive murine pancreatic centroacinar and terminal duct cells was confirmed.

Conclusions:

AldeRed 588-A is a valuable tool for selecting stem cells based on ALDH activity.
This reagent facilitates the study of ALDH in biological systems, especially when green fluorescent reporters are present.
The development of AldeRed 588-A overcomes limitations of existing reagents for ALDH-positive cell identification.