Isolation of primitive human hematopoietic progenitors on the basis of aldehyde dehydrogenase activity.
Because hematopoietic stem cells are rich in aldehyde dehydrogenase (ALDH) activity, we developed a fluorescent substrate for ALDH, termed BODIPY aminoacetaldehyde (BAAA), and tested its potential for isolating primitive human hematopoietic cells. A population of cells with low orthogonal light scattering and bright fluorescence intensity (SSC(lo)ALDH(br) cells) could be readily fractionated from human umbilical cord blood cells costained with BAAA and the multidrug-resistance inhibitor verapamil. The SSC(lo)ALDH(br) population was depleted of lineage-committed cells, 40-90% pure for CD34(+)CD38(lo/-) cells, and enriched 50- to 100-fold for primitive hematopoietic progenitors detected in short- and long-term culture analyses. Together, these observations indicate that fractionating human hematopoietic stem cells on the basis of ALDH activity using BAAA is an effective method for isolating primitive human hematopoietic progenitors. This technique may be useful for isolating stem cells from other tissues as well.
Storms
R W
RW
The Center for Genetic and Cellular Therapies, Department of Experimental Surgery, Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
Trujillo
A P
AP
Springer
J B
JB
Shah
L
L
Colvin
O M
OM
Ludeman
S M
SM
Smith
C
C
D000444:Aldehyde Dehydrogenase / Q000378:metabolism*; D001896:Boron Compounds / Q000138:chemical synthesis; D002469:Cell Separation / Q000379:methods*; D003114:Colony-Forming Units Assay; D004396:Coloring Agents; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000201:enzymology*; D006801:Humans; D007231:Infant, Newborn; D020014:K562 Cells; D013379:Substrate Specificity; D014700:Verapamil
C120929:BODIPY acetoacetaldehyde; D001896:Boron Compounds; D004396:Coloring Agents; D014700:Verapamil; D000444:Aldehyde Dehydrogenase