ALDEFLUOR™

Identification, Evaluation and Isolation of Normal and Cancer Stem & Progenitor Cells

ALDEFLUOR™ is a non-immunological fluorescent reagent system that has supported over 1000 publications for the detection of aldehyde dehydrogenase-bright (ALDHbr) cells in over 80 different tissues. High expression of ALDH has been reported for normal and cancer stem and progenitor cells of various lineages, including hematopoietic, mammary, endothelial, mesenchymal and neural cells. Only cells with an intact cellular membrane can retain the ALDEFLUOR™ reaction product, making this system selective for viable ALDHbr cells. ALDEFLUOR™ is a non-toxic and easy-to-use kit that requires no antibody staining, and is compatible with standard cell sorters and analyzers.

How ALDEFLUOR™ Detects Normal and Cancer Progenitor Cells

The ALDEFLUOR™ kit contains BODIPY-aminoacetaldehyde (BAAA), a fluorescent non-toxic substrate for ALDH, which freely diffuses into intact and viable cells. In the presence of ALDH, BAAA is converted into BODIPY-aminoacetate (BAA), a negatively charged product that is retained inside the cells. Intracellular accumulation of BAA leads to increased fluorescence, and these ALDH-bright (ALDHbr) cells can be analyzed with flow cytometry. Watch this video to learn more about how ALDEFLUOR™ works.

Optimizing ALDEFLUOR™ for Various Tissue Types

ALDEFLUOR™ was initially developed and optimized for the detection of ALDHbr hematopoietic stem and progenitor cells in human blood and bone marrow. Since then it has also been shown to detect normal and neoplastic cells in many other tissue types (including breast, colon, lung, pancreas and thyroid) as well as cancer cell lines. Optimizing the ALDEFLUOR™ protocol to your tissue type of interest can dramatically increase the fluorescence intensity, thereby optimizing the assay performance. Watch this video to see an example of how our scientists increased the fluorescence intensity of ALDHbr cells in mammary tissue, and how you can modify the protocol to fit your research needs.

Why Use ALDEFLUOR™?

  • Detect viable normal or cancer progenitor cells based on ALDH activity. No antibody staining required.
  • Can be used with multiple cell types and species.
  • Identifies only viable cells with an intact cell membrane. Compatible with immunophenotyping.
  • Has supported more than 1000 peer-reviewed publications.
  • Simple protocol with highly reproducible results. Compatible with standard cell sorters or analyzers.

ALDEFLUOR™ Products

ALDEFLUOR™ Kit

ALDEFLUOR™ Kit

Recommended for:

Detection of stem and progenitor cells from normal and cancerous tissue

Species:

Human, Mouse, Rat

ALDHbr Assay Kit

ALDEFLUOR™ Kit

Recommended for:

Detection of ALDHbrCD34+ hematopoietic stem and progenitor cells in cord blood

Species:

Human

ALDEFLUOR™ Assay Buffer

ALDEFLUOR™ Kit

Recommended for:

Counterstaining ALDEFLUOR™-reacted cells with immunofluorescently-labeled antibodies

Species:

Human, Mouse, Rat
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Key Applications

Publications on Normal Cells

Hematopoietic Cells

Boxall SA et al. (2008) Haematopoietic repopulating activity in human cord blood CD133+ quiescent cells. Bone Marrow Transplant 43(8): 627-35.
Capoccia BJ et al. (2009) Revascularization of ischemic limbs after transplantation of human bone marrow cells with high aldehyde dehydrogenase activity. Blood 113(21): 5340-51.
Christ O et al. (2007) Improved purification of hematopoietic stem cells based on their elevated aldehyde dehydrogenase activity. Haematologica 92(9): 1165-72.
Fallon P et al. (2003) Mobilized peripheral blood SSCloALDHbr cells have the phenotypic and functional properties of primitive haematopoietic cells and their number correlates with engraftment following autologous transplantation. Br J Haematol 122: 99-108.
Gentry T et al. (2007) Isolation of early hematopoietic cells, including megakaryocyte progenitors, in the ALDH-bright cell population of cryopreserved, banked UC blood. Cytotherapy 9(6): 569-76.
Gentry T et al. (2007) Simultaneous isolation of human BM hematopoietic, endothelial and mesenchymal progenitor cells by flow sorting based on aldehyde dehydrogenase activity: implications for cell therapy. Cytotherapy 9(3): 259-74.
Gündüz E et al. (2010) Evaluation of mobilized peripheral stem cells according to CD34 and aldehyde dehydrogenase expression and effect of SSC(lo) ALDH(br) cells on hematopoietic recovery. Cytotherapy 12(8): 1006-12.
Hess DA et al. (2004) Functional characterization of highly purified human hematopoietic repopulating cells isolated according to aldehyde dehydrogenase activity. Blood 104(6): 1648-55.
Hess DA et al. (2008) Widespread nonhematopoietic tissue distribution by transplanted human progenitor cells with high aldehyde dehydrogenase activity. Stem Cells 26(3): 611-20.
Liu C et al. (2010) Progenitor cell dose determines the pace and completeness of engraftment in a xenograft model for cord blood transplantation. Blood 116(25): 5518-27.
Muramoto GG et al. (2010) Inhibition of aldehyde dehydrogenase expands hematopoietic stem cells with radioprotective capacity. Stem Cells 28(3): 523-34.
Pearce DJ & Connet D. (2007) The combined use of Hoechst efflux ability and aldehyde dehydrogenase activity to identify murine and human hematopoietic stem cells. Exp Hematol 35(9): 1437-46.
Pierre-Louis O et al. (2009) Dual SP/ALDH functionalities refine the human hematopoietic Lin-CD34+CD38- stem/progenitor cell compartment. Stem Cells 27(10): 2552-62.
Povsic TJ et al. (2009) Aldehyde dehydrogenase activity allows reliable EPC enumeration in stored peripheral blood samples. J Thromb Thrombolysis 28(3): 259-65.
Povsic TJ et al. (2010) Aging is not associated with bone marrow-resident progenitor cell depletion. J Gerontol A Biol Sci Med Sci 65(10): 1042-50.
Sondergaard CS et al. (2010) Human cord blood progenitors with high aldehyde dehydrogenase activity improve vascular density in a model of acute myocardial infarction. J Transl Med 8: 24.
Storms RW et al. (1999) Isolation of primitive human hematopoietic progenitors on the basis of aldehyde dehydrogenase activity. Proc Natl Acad Sci U S A 96: 9118-23.
Shoulars K et al. (2016) Development and validation of a rapid, aldehyde dehydrogenase bright-based cord blood potency assay. Blood 127(19):2346-54.

Endothelial Cells

Mammary Cells

Ginestier C et al. (2007) ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell 1(5): 555-67.
Liu S et al. (2008) BRCA1 regulates human mammary stem/progenitor cell fate. Proc Natl Acad Sci U S A 105(5): 1680-5.

Mesenchymal Cells

Pancreatic Cells

Prostate Cells

Yao M et al. (2010) Prostate-regenerating capacity of cultured human adult prostate epithelial cells. Cells Tissues Organs 191(3): 203-12.

Publications on Cancer Cells

Cancer Stem Cells: Review Articles

Alison MR et al. (2011) Cancer Stem Cells: Problems for Therapy? J Pathol 223(2): 147-161.
Alison MR et al. (2010) Finding Cancer Stem Cells: Are Aldehyde Dehydrogenases Fit for Purpose? J Pathol 222(4): 335-44.
Ma I & Allan AL. (2011) The Role of Human Aldehyde Dehydrogenase in Normal and Cancer Stem Cells. Stem Cell Rev & Rep 7(2): 292-306.

Breast Cancer Cells

Alam M et al. (2013) MUC1-C Oncoprotein Activates ERK→C/EBP Signaling and Induction of Aldehyde Dehydrogenase 1A1 in Breast Cancer Cells. J Biol Chem 288(43): 30892-903.
Atkinson RL et al. (2013) Cancer Stem Cell Markers Are Enriched in Normal Tissue Adjacent to Triple Negative Breast Cancer and Inversely Correlated with DNA Repair Deficiency. Breast Cancer Res 15(5): R77.
Azzam DJ et al. (2013) Triple Negative Breast Cancer Initiating Cell Subsets Differ in Functional and Molecular Characteristics and in γ-Secretase Inhibitor Drug Responses. EMBO Mol Med 5(10): 1502-22.
Buckley NE et al. (2013) BRCA1 Is a Key Regulator of Breast Differentiation Through Activation of Notch Signalling with Implications for Anti-Endocrine Treatment of Breast Cancers. Nucl Acids Res 41(18): 8601-14.
Buijs JT et al. (2012) The BMP2/7 Heterodimer Inhibits the Human Breast Cancer Stem Cell Subpopulation and Bone Metastases Formation. Oncogene 31(17): 2164-74.
Chen D et al. (2013) ANTXR1, a Stem Cell-Enriched Functional Biomarker, Connects Collagen Signaling to Cancer Stem-Like Cells and Metastasis in Breast Cancer. Cancer Res 73(18): 5821-33.
Conti L et al. (2013) The Noninflammatory Role of High Mobility Group Box 1/Toll-Like Receptor 2 Axis in the Self-Renewal of Mammary Cancer Stem Cells. FASEB J 27(12): 4731-44.
Ithimakin S et al. (2013) HER2 Drives Luminal Breast Cancer Stem Cells in the Absence of HER2 Amplification: Implications for Efficacy of Adjuvant Trastuzumab. Cancer Res 73(5): 1635-46.
Kundu N et al. (2014) Prostaglandin E Receptor EP4 Is a Therapeutic Target in Breast Cancer Cells with Stem-Like Properties. Breast Cancer Res TR 143(1): 19-31.
Liu S et al. (2008) BRCA1 Regulates Human Mammary Stem/ Progenitor Cell Fate. PNAS 105(5): 1680-85.
Liu P et al. (2013) Disulfiram Targets Cancer Stem-Like Cells and Reverses Resistance and Cross-Resistance in Acquired Paclitaxel-Resistant Triple-Negative Breast Cancer Cells. Brit J Cancer 109(7): 1876-85.
Londoño-Joshi AI et al. (2014) Effect of Niclosamide on Basal-Like Breast Cancers. Mol Cancer Ther 13(4):800-11.
McClements L et al. (2013) Targeting Treatment-Resistant Breast Cancer Stem Cells with FKBPL and its Peptide Derivative, AD-01, Via the CD44 Pathway. Clin Cancer Res 19(14): 3881-93.
Piva M et al. (2014) Sox2 Promotes Tamoxifen Resistance in Breast Cancer Cells. EMBO Mol Med 6(1): 66-79.
Rustighi A et al. (2014) Prolyl-Isomerase Pin1 Controls Normal and Cancer Stem Cells of the Breast. EMBO Mol Med 6(1): 99-119.
Salvador MA et al. (2013) The Histone Deacetylase Inhibitor Abexinostat Induces Cancer Stem Cells Differentiation in Breast Cancer with Low Xist Expression. Clin Cancer Res 19(23): 6520-31.
Vazquez-Martin A et al. (2013) Reprogramming of Non-Genomic Estrogen Signaling by the Stemness Factor SOX2 Enhances the Tumor-Initiating Capacity of Breast Cancer Cells. Cell Cycle 12(22): 3471-77.
Wang X et al. (2013) PPARγ Maintains ERBB2-Positive Breast Cancer Stem Cells. Oncogene 32(49): 5512-21.
Yamamoto M et al. (2013) NF-κB Non-Cell-Autonomously Regulates Cancer Stem Cell Populations in the Basal-Like Breast Cancer Subtype. Nat Commun 4: 2299.
Yu F et al. (2011) Kruppel-Like Factor 4 (KLF4) is Required for Maintenance of Breast Cancer Stem Cells and for Cell Migration and Invasion. Oncogene 30(18): 2161-272.
Zhou Y et al. (2014) The miR-106b25 Cluster Promotes Bypass of Doxorubicin-Induced Senescence and Increase in Motility and Invasion by Targeting the E-Cadherin Transcriptional Activator EP300. Cell Death Differ 21(3): 462-74.

Skin Cancer Cells

Boonyaratanakornkit JB et al. (2010) Selection of Tumorigenic Melanoma Cells Using ALDH. J Invest Dermatol 130(12): 2799– 808.

Thyroid Cancer Cells

Todaro M et al. (2010) Tumorigenic and Metastatic Activity of Human Thyroid Cancer Stem Cells. Cancer Res 70(21): 8874-85.
ALDEFLUOR is a trademark of Aldagen Inc.
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