Determination of mononuclear cell count using peripheral smear and flow cytometry in peripheral blood stem cell products

Abstract

BACKGROUND: Mononuclear cells (MNCs) are considered equivalent to hematopoietic stem cells, and differential count using peripheral smear was routinely practiced to enumerate MNC. Flow cytometry plots used for CD34 enumeration assay can also be used in MNC enumeration as it counts more WBC events than manual methods. The aim was to determine the relationship and degree of agreement between peripheral smear and flow cytometry in MNC enumeration of peripheral blood stem cell (PBSC) products. METHODS: In 63 patients, 73 PBSC products were collected between January 2017 and September 2019. The differences in MNC count estimated by peripheral smear method and from flow cytometry plots used for CD34 enumeration were analyzed using Mann–Whitney test. Agreement between the two methods for MNC enumeration was determined by regression analysis. Receiver operating characteristic curve was performed to determine MNC threshold in peripheral blood and PBSC product for adequate mobilization and harvest. RESULTS: There was no difference in enumeration of median MNC count between peripheral smear and flow cytometry (52% vs. 59%, P = 0.185) in PBSC product. However, regression analysis indicated a constant and proportional difference between the methods with r = 0.52. Cumulative sum test for linearity showed deviation from linearity ( P = 0.04). MNC counts in peripheral blood failed to achieve discrimination capacity in predicting adequate CD34+ yield/kg body weight in product. CONCLUSION: Peripheral smear estimated lower MNC counts than flow cytometry with weaker agreements between the two methods. Hence, MNC count derived from flow cytometry plot can substitute peripheral smear method for MNC dose calculations. MNC dose at 3.4 × 10 8 /kg consistently predicted >2 × 10 6 /kg CD34+ cells collected.