Limits...
Next generation of ALDH substrates and their potential to study maturational lineage biology in stem and progenitor cells.

Dollé L, Boulter L, Leclercq IA, van Grunsven LA - Am. J. Physiol. Gastrointest. Liver Physiol. (2015)

Bottom Line: In this perspective, novel ALDH substrates are being developed.Here we describe how new substrates could be instrumental for better isolation of cell population with stemness potential and for defining hierarchy of cell populations in tissue.Finally, we speculate on other potential applications.

View Article: PubMed Central - PubMed

Affiliation: Liver Cell Biology Lab, Vrije Universiteit Brussel (VUB), Brussels, Belgium; ldolle@vub.ac.be.

Show MeSH

Related in: MedlinePlus

The AldeRed-588-A substrate provides a useful tool to select stem cells and opens up prospects for the next generation of aldehyde dehydrogenase (ALDH) substrates. A: Aldefluor and AldeRed-588-A have the same capacity to isolate an ALDHbright cell population enriched in stem cells from a heterogeneous mixture of cells. B: AldeRed-588-A can be used for multicolor applications to fractionate ALDHbright cells in the presence of green fluorophores, including the Aldefluor reagent and cells expressing enhanced green-fluorescent protein (eGFP) and can be visualized by fluorescent microscopy. C: emerging opportunities in generating preferred labeled substrates with different fluorescent probes. D: distinguishing distinct cell populations from a digested tissue on the basis of the specificity of the ALDH substrate might help in fractionating the different category of cells with ALDH activity referring to a certain physiological state. ALDHbright, cells with high ALDH activity; ALDHdim, low-ALDH-activity fraction; FSC, forward-scattered light; SSC, side-scattered light.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4385895&req=5

Figure 1: The AldeRed-588-A substrate provides a useful tool to select stem cells and opens up prospects for the next generation of aldehyde dehydrogenase (ALDH) substrates. A: Aldefluor and AldeRed-588-A have the same capacity to isolate an ALDHbright cell population enriched in stem cells from a heterogeneous mixture of cells. B: AldeRed-588-A can be used for multicolor applications to fractionate ALDHbright cells in the presence of green fluorophores, including the Aldefluor reagent and cells expressing enhanced green-fluorescent protein (eGFP) and can be visualized by fluorescent microscopy. C: emerging opportunities in generating preferred labeled substrates with different fluorescent probes. D: distinguishing distinct cell populations from a digested tissue on the basis of the specificity of the ALDH substrate might help in fractionating the different category of cells with ALDH activity referring to a certain physiological state. ALDHbright, cells with high ALDH activity; ALDHdim, low-ALDH-activity fraction; FSC, forward-scattered light; SSC, side-scattered light.

Mentions: By their recent publication in Nature Communications, Minn and collaborators (21) rejuvenated the interest in using ALDH activity. They described a new red-shifted fluorescent ALDH substrate (AldeRed-588-A), for labeling of viable ALDHbright cell populations. Structurally, Aldefluor (or Bodipy493/503-aminoacetaldehyde) and AldeRed-588-A (or Bodipy576/589-aminoacetaldehyde) have a common substrate moiety: acetaldehyde, indicating that a cell oxidizing Aldefluor is likely to have the same ability to metabolize the AldeRed-588-A substrate (Fig. 1A). In other words, AldeRed-588-A is considered as one of the other alternatives to Aldefluor substrate, without other functionally different characteristics. Elegantly, using cell lines known to express functional ALDH the authors demonstrated that Aldefluor and AldeRed-588-A essentially have the same efficacy and efficiency for identifying a ALDHbright population (21). In addition, by successfully mixing two substrates, Minn and colleagues prove that labeling technique does not impede the structural recognition of the substrate by ALDH enzyme and that cell isolation of ALDH expressing cells is feasible by a single-step isolation method (Aldefluor and AldeRed-588-A are incubated simultaneously), thus avoiding additional purification or enrichment steps in which cells can be lost or damaged. This new AldeRed-588-A substrate has a broad emission spectrum, which currently precludes combination with a large array of fluorophores. Nevertheless, the technical innovation opens new avenues for stem cell research by offering a greater flexibility for ALDHbright cell isolations.


Next generation of ALDH substrates and their potential to study maturational lineage biology in stem and progenitor cells.

Dollé L, Boulter L, Leclercq IA, van Grunsven LA - Am. J. Physiol. Gastrointest. Liver Physiol. (2015)

The AldeRed-588-A substrate provides a useful tool to select stem cells and opens up prospects for the next generation of aldehyde dehydrogenase (ALDH) substrates. A: Aldefluor and AldeRed-588-A have the same capacity to isolate an ALDHbright cell population enriched in stem cells from a heterogeneous mixture of cells. B: AldeRed-588-A can be used for multicolor applications to fractionate ALDHbright cells in the presence of green fluorophores, including the Aldefluor reagent and cells expressing enhanced green-fluorescent protein (eGFP) and can be visualized by fluorescent microscopy. C: emerging opportunities in generating preferred labeled substrates with different fluorescent probes. D: distinguishing distinct cell populations from a digested tissue on the basis of the specificity of the ALDH substrate might help in fractionating the different category of cells with ALDH activity referring to a certain physiological state. ALDHbright, cells with high ALDH activity; ALDHdim, low-ALDH-activity fraction; FSC, forward-scattered light; SSC, side-scattered light.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4385895&req=5

Figure 1: The AldeRed-588-A substrate provides a useful tool to select stem cells and opens up prospects for the next generation of aldehyde dehydrogenase (ALDH) substrates. A: Aldefluor and AldeRed-588-A have the same capacity to isolate an ALDHbright cell population enriched in stem cells from a heterogeneous mixture of cells. B: AldeRed-588-A can be used for multicolor applications to fractionate ALDHbright cells in the presence of green fluorophores, including the Aldefluor reagent and cells expressing enhanced green-fluorescent protein (eGFP) and can be visualized by fluorescent microscopy. C: emerging opportunities in generating preferred labeled substrates with different fluorescent probes. D: distinguishing distinct cell populations from a digested tissue on the basis of the specificity of the ALDH substrate might help in fractionating the different category of cells with ALDH activity referring to a certain physiological state. ALDHbright, cells with high ALDH activity; ALDHdim, low-ALDH-activity fraction; FSC, forward-scattered light; SSC, side-scattered light.
Mentions: By their recent publication in Nature Communications, Minn and collaborators (21) rejuvenated the interest in using ALDH activity. They described a new red-shifted fluorescent ALDH substrate (AldeRed-588-A), for labeling of viable ALDHbright cell populations. Structurally, Aldefluor (or Bodipy493/503-aminoacetaldehyde) and AldeRed-588-A (or Bodipy576/589-aminoacetaldehyde) have a common substrate moiety: acetaldehyde, indicating that a cell oxidizing Aldefluor is likely to have the same ability to metabolize the AldeRed-588-A substrate (Fig. 1A). In other words, AldeRed-588-A is considered as one of the other alternatives to Aldefluor substrate, without other functionally different characteristics. Elegantly, using cell lines known to express functional ALDH the authors demonstrated that Aldefluor and AldeRed-588-A essentially have the same efficacy and efficiency for identifying a ALDHbright population (21). In addition, by successfully mixing two substrates, Minn and colleagues prove that labeling technique does not impede the structural recognition of the substrate by ALDH enzyme and that cell isolation of ALDH expressing cells is feasible by a single-step isolation method (Aldefluor and AldeRed-588-A are incubated simultaneously), thus avoiding additional purification or enrichment steps in which cells can be lost or damaged. This new AldeRed-588-A substrate has a broad emission spectrum, which currently precludes combination with a large array of fluorophores. Nevertheless, the technical innovation opens new avenues for stem cell research by offering a greater flexibility for ALDHbright cell isolations.

Bottom Line: In this perspective, novel ALDH substrates are being developed.Here we describe how new substrates could be instrumental for better isolation of cell population with stemness potential and for defining hierarchy of cell populations in tissue.Finally, we speculate on other potential applications.

View Article: PubMed Central - PubMed

Affiliation: Liver Cell Biology Lab, Vrije Universiteit Brussel (VUB), Brussels, Belgium; ldolle@vub.ac.be.

Show MeSH
Related in: MedlinePlus