Abstract
Aldehyde dehydrogenase (ALDH) is a cytosolic enzyme that is responsible for the oxidation of intracellular aldehydes. Elevated levels of ALDH have been demonstrated in murine and human progenitor cells compared with other hematopoietic cells, and this is thought to be important in chemoresistance and purification techniques and an indication of the proper function of the cell. A Flowcytometric method for the assessment of ALDH activity in viable cells recently has been developed. Forty six cord blood samples from mothers which underwent normal delivery of full term infants were obtained, after informed consent. Mononuclear cells were obtained by Ficoll-Paque density centrifugation and ammonium chloride red cell lysis. Percentage of viable cells was determined by trypan blue exclusion dye. Cells were labeled with Aldefluor reagent (Vancouver Canada) as described by the manufacturer. Cells were then stained with phycoerythrin (PE)–conjugated anti-CD34 (Miltenyi Biotec, Cologne, Germany) antibodies for 30 min at 4°C. Cells were washed and re-suspended in phosphate-buffered saline (PBS) with 2% fetal calf serum. Cells were then analyzed on coulter epics flow cytometer. The mean percentage of ALDH enzyme expression among the CD34+ cells in the cord blood samples was 61.3% with a minimum of 28% and a maximum of 94.6%. Significant correlations were found between the white blood cell (WBCS) count in the cord blood samples and both the CD34+ cell count and the count of ALDH expressing cells, while no correlation was found between the CD34+ cells count or the ALDH expressing cells count in the cord blood samples and either the sex or the weight of the newborn. Identification and isolation of cells on the basis of ALDH activity provides a tool for their isolation and further analysis. In summary, a high ALDH-1 activity identifies CD34+ cells in cord blood. Key words: Umbilical cord blood, stem cells, aldehyde dehydrogenase (ALDH), CD34.
Highlights
Aldehyde dehydrogenase (ALDH) is a family of enzymes involved in metabolism of aldehydes to their corresponding carboxylic acids (Cheung et al, 2007)
The study population included 46 umbilical cord blood samples collected from immediate newborns, 22 males (47.8%) and 24 females (52.2%)
We found a statistically significant difference when we compared between the mean count of CD34+ cells and the mean count of ALDH expressing cells (Table 3)
Summary
Aldehyde dehydrogenase (ALDH) is a family of enzymes involved in metabolism of aldehydes to their corresponding carboxylic acids (Cheung et al, 2007) It plays an important role in metabolism of vitamin A as well as in mechanisms of resistance to alkylating agents, for example cyclophosphamide (Storms et al, 1999). UCB cells isolated by using this strategy have demonstrated to be depleted of lineage committed hematopoietic cells and are enriched for primitive hematopoietic progenitors detected in clonogenic in vitro cultures (Storms et al, 1999) This approach has allowed the analysis of viable murine and human ALDH+ progenitors by flow cytometry (Storms et al, 1999)
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