Assessment of aldehyde dehydrogenase in viable cells

Richard J. Jones, James P. Barber, Milada S. Vala, Michael I. Collector, Scott H Kaufmann, Susan M. Ludeman, O. Michael Colvin, John Hilton

Research output: Contribution to journalArticle

240 Citations (Scopus)

Abstract

Cytosolic aldehyde dehydrogenase (ALDH), an enzyme responsible for oxidizing intracellular aldehydes, has an important role in ethanol, vitamin A, and cyclophosphamide metabolism. High expression of this enzyme in primitive stem cells from multiple tissues, including bone marrow and intestine, appears to be an important mechanism by which these cells are resistant to cyclophosphamide. However, although hematopoietic stem cells (HSC) express high levels of cytosolic ALDH, isolating viable HSC by their ALDH expression has not been possible because ALDH is an intracellular protein. We found that a fluorescent aldehyde, dansyl aminoacetaldehyde (DAAA), could be used in flow cytometry experiments to isolate viable mouse and human cells based on their ALDH content. The level of dansyl fluorescence exhibited by cells after incubation with DAAA paralleled cytosolic ALDH levels determined by Western blotting and the sensitivity of the cells to cyclophosphamide. Moreover, DAAA appeared to be a more sensitive means of assessing cytosolic ALDH levels than Western blotting. Bone marrow progenitors treated with DAAA proliferated normally. Furthermore, marrow cells expressing high levels of dansyl fluorescence after incubation with DAAA were enriched for hematopoietic progenitors. The ability to isolate viable cells that express high levels of cytosolic ALDH could be an important component of methodology for identifying and purifying HSC and for studying cyclophosphamide-resistant tumor cell populations.

Original languageEnglish (US)
Pages (from-to)2742-2746
Number of pages5
JournalBlood
Volume85
Issue number10
StatePublished - May 15 1995
Externally publishedYes

Fingerprint

Aldehyde Dehydrogenase
Stem cells
Cyclophosphamide
Hematopoietic Stem Cells
Bone Marrow
Cells
Aldehydes
Bone
Fluorescence
Western Blotting
Flow cytometry
Enzymes
Vitamin A
Metabolism
Intestines
Tumors
Flow Cytometry
Ethanol
Stem Cells
Tissue

ASJC Scopus subject areas

  • Hematology

Cite this

Jones, R. J., Barber, J. P., Vala, M. S., Collector, M. I., Kaufmann, S. H., Ludeman, S. M., ... Hilton, J. (1995). Assessment of aldehyde dehydrogenase in viable cells. Blood, 85(10), 2742-2746.

Assessment of aldehyde dehydrogenase in viable cells. / Jones, Richard J.; Barber, James P.; Vala, Milada S.; Collector, Michael I.; Kaufmann, Scott H; Ludeman, Susan M.; Colvin, O. Michael; Hilton, John.

In: Blood, Vol. 85, No. 10, 15.05.1995, p. 2742-2746.

Research output: Contribution to journalArticle

Jones, RJ, Barber, JP, Vala, MS, Collector, MI, Kaufmann, SH, Ludeman, SM, Colvin, OM & Hilton, J 1995, 'Assessment of aldehyde dehydrogenase in viable cells', Blood, vol. 85, no. 10, pp. 2742-2746.
Jones RJ, Barber JP, Vala MS, Collector MI, Kaufmann SH, Ludeman SM et al. Assessment of aldehyde dehydrogenase in viable cells. Blood. 1995 May 15;85(10):2742-2746.
Jones, Richard J. ; Barber, James P. ; Vala, Milada S. ; Collector, Michael I. ; Kaufmann, Scott H ; Ludeman, Susan M. ; Colvin, O. Michael ; Hilton, John. / Assessment of aldehyde dehydrogenase in viable cells. In: Blood. 1995 ; Vol. 85, No. 10. pp. 2742-2746.
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