ISSN: 1300-7777 E-ISSN: 1308-5263
Flow Cytometrıc Aldehyde Dehydrogenase (Aldh) Assay Enables A Fast And Accurate Human Umblical Cord Blood Hematopoietic Stem Cell Assessment [Turk J Hematol]
Turk J Hematol. Ahead of Print: TJH-66743 | DOI: 10.4274/tjh.2016.0214  

Flow Cytometrıc Aldehyde Dehydrogenase (Aldh) Assay Enables A Fast And Accurate Human Umblical Cord Blood Hematopoietic Stem Cell Assessment

Emine Begüm Gencer1, Pınar Yurdakul2, Klara Dalva3, Meral Beksac4
1Cord Blood Bank, Ankara University School of Medicine, Ankara, Turkey, Biotechnology Institute, Ankara University, Ankara, Turkey
2Cord Blood Bank, Ankara University School of Medicine, Ankara, Turkey, Department of Medical Microbiology, TOBB ETU School of Medicine, Ankara, Turkey
3Stem Cell Research Institute, Ankara University School of Medicine, Ankara, Turkey
4Department of Hematology, Ankara University School of Medicine, Ankara, Turkey

Objective: Colony forming unit-granulocyte macrophage (CFU-GM) analysis is the most widely used method to determine hematopoieticstem cell (HSC) content of human umbilical cord blood (CB) for prediction of engraftment potential. Measurement of aldehyde dehydrogenase (ALDH) activity is a more recent method for HSC qualification. Our aim was to correlate phenotypic with functional assays to find the most predictive method.

Materials and Methods: In this study, flow cytometric quantitation of CD34+ cells and ALDH positivity along with CFU-GM capacity were assessed in fresh and post thaw CB units.
Results: Among 30 post-process samples: For each CB unit the mean total number of nucleated cells (TNC): 93,8 ± 30,1 x107, CD34+: 3,85 ± 2,55 x106, ALDH+: 3,14 ± 2,55 x106, CFU-GM: 2,64 ± 1,96 x105. Among additional 19 post thaw samples the cell counts were: TNC: 32,79 ± 17,27 x107, CD34+: 2,18 ± 3,17 x106, ALDH+ 2,01 ± 2,81 x106, CFU-GM: 0,74 ± 0,92 x105. Our findings showed that, in fresh samples, TNC, CD34 cells and ALDH correlated highly with CFU-GM, CFU-GEMM and BFU-E as such: TNC, r=0,47, r=0,35, r=0,41; CD34+, r=0,44, r=0,54 r=0,41; and ALDH, r=0,63 r=0,45 r=0,6, respectively. In terms of post thaw samples, the correlations were: TNC r=0,59, r=0,46, r=0,56, CD34+ r=0,67, r=0,48, r=0,61 and ALDH r=0,61, r=0,67, r=0,67 for CFU-GM, CFU-GEMM, and BFU-E, respectively. All correlations were statistically significant.
Conclusion: In our experience, HSC assessment by ALDH activity yields highest correlation with conventional analytic methods, particularly among post-thaw samples. Thus this fast, inexpensive method has the potential to overcome the weaknesses of these techniques.

Keywords: Cord blood, ALDH, CFU-GM




Corresponding Author: Meral Beksac, Türkiye


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