Cristian Nunez-Espinosa, María D. Garcia-Godoy, Ines Ferreira, Juan G. Rios-Kristjansson, David Rizo-Roca, Laura G. Rico, Gerard Rubi-Sans, Carlos Palacio, Joan R. Torrella, Teresa Pages, Mike D. Ward, Gines Viscor and Jordi Petriz Pages 66 - 71 ( 6 )
Introduction: Studies are needed to understand the role of CD34 expressing cells with regard to efficient engraftment, especially in the adjuvant treatment of cancer. Materials and Methods: In this study we have used a modified method in our laboratory for routinely counting CD34+ cells. Unlysed whole blood samples were stained with the DNA-selective and cell membrane-permeant Vibrant DyeCycle Violet stain. Results: CD34+ cells exhibit a consistent and differential Vybrant Dye Cycle Violet staining pattern. Based on their different DCV intensity, we classified these subpopulations as CD34+/DCVhigh and CD34+/DCVlow cells. In general, DCVhigh cells are about 12-times brighter than DCVlow cells. Conclusion: DCV staining may be used to discriminate subsets of CD34+ cells similarly to other methods which have previously defined different functional properties that can be related to the characterization, resolution, and purification of primitive hematopoietic stem cells in combination with specific useful markers for multicolor flow cytometric measurements.
CD34, flow cytometry, P-glycoprotein, Stem cells, Vybrant DyeCycle Violet.
Josep Carreras Leukemia Research Institute, Crta. de Can Ruti, Camí de les Escoles s/n. Edifici IMPPC, 08916 Badalona (Barcelona), Spain.