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Comparison of CD34+ cell enumeration between flow cytometric analysis and ADAMII-CD34 image-based fluorescence cell
Anchalee Thedsawad1, Sirikwan Boonmoh1, Orathai Taka1
1Division of Hematology, Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok Noi, Bangkok, Thailand.
Background:
Accurate CD34+ cell enumeration is essential in stem cell transplantation. Flow cytometry (FC) is the standard method to determine optimal collection time of CD34+ stem cells harvesting for peripheral blood stem cell (PBSC) transplantation but is costly and requires skilled personnel. The ADAMII-CD34 cell counter may be a feasible alternative for PBSC apheresis samples and for cryopreserved stem cell products which are assessed by manual cell viability counting using the trypan blue exclusion method.
Methods:
Fifty samples were analyzed (PBSC apheresis samples n = 25, cryopreserved stem cell products n = 25) from patients undergoing autologous PBSC transplantation or healthy donors. We assessed the quantitative agreement between methods.
Results:
In the 25 PBSC apheresis samples, FC-based median viable CD34+ absolute count was 764.7 cells/µL (IQR 453.7-1532.5; range 60.5-3233.8). ADAMII-CD34 cell counter and FC showed almost perfect agreement [Lin's concordance correlation coefficient (CCC) 0.99 (95% CI: 0.98 to 1.00), bias -0.46% (95% CI: -5.45 to 4.53), upper limit of agreement (LoA) 22.2% (95% CI: 13.5 to 30.8), and lower LoA -23.1% (95% CI: -31.7 to -14.4)]. For cryopreserved stem cell products, viable CD34+ cell dose (×106 cells/kg) measured by the ADAMII-CD34 cell counter and the estimated viable CD34+ cell dose (×106 cells/kg) determined by the trypan blue exclusion method showed poor agreement [Lin's CCC 0.58 (95% CI: 0.31 to 0.76)]. Weighted Deming regression showed proportional bias [95% CI: slope 0.02 to 0.61].
Conclusions:
Quantitative agreement between the ADAMII-CD34 cell counter and FC for CD34+ enumeration in PBSC apheresis samples was acceptable within the analyzed range. It is user-friendly and more affordable than FC. However, ADAMII-CD34 and the trypan blue exclusion method of cryopreserved stem cell products showed a proportional bias. Further considerations are needed to optimize the process for cryopreserved stem cell products.
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