Prospective flow cytometric evaluation of nucleated red blood cells in cord blood units and relationship with nucleated and CD34+ cell quantification

BACKGROUND:  Cord blood (CB) represents an alternate source of stem cells in transplantation. Nucleated red blood cells (NRBCs) are a physiological subset of CB population. Although it is important to have an accurate estimate of CD34+ cell number, NRBCs could compromise white blood cell count and interfere with CD34+ cell quantification. STUDY DESIGN AND METHODS:  A total of 826 CB units were analyzed for total nucleated cells (TNCs), NRBCs, and CD34+ cells by flow cytometry. NRBCs were also counted conventionally by manual microscopy. Percentages of CD34+ cells corrected by NRBC count (CD34+c) were determined as follows: %CD34+c = CD34+/CD45+ (×10 6 )/(TNCs (×10 8 ) – NRBCs (×10 8 )). RESULTS:  The mean percentages of CD34+ cells and NRBCs were 0.27 percent (range, 0.01%‐1.25%) and 7.64 percent (range, 0.13%‐84%), respectively. Comparison between flow cytometric and microscopic NRBC count showed a regression of y = 0.685 + 0.719x and a coefficient of determination of r 2  = 0.721. When corrected with NRBC count, the mean percentage of CD34+ c cells was 0.295 percent (p = 0.0008 compared with CD34+%) and mean TNCc count was 14.8 × 10 8 (p < 10 −4 compared to TNC count). CONCLUSION:  The determination of NRBCs with a flow cytometric method might represent a new strategy for providing satisfactory quality assurance controls of CB products.