An internal positive control for the enumeration of CD45+ and CD34+ cells by flow cytometry allows monitoring of reagent and operator performance

Human bone marrow processing using a new continuous‐flow cell separation device

“…The WBC count, Hct, and MNC count were determined using a hematology analyzer (LH750, Beckman Coulter, High Wycombe, Buckinghamshire, UK). The CD45+, CD34+, and CD3+ cell counts were obtained by a single‐platform, three‐color flow cytometry assay using a flow cytometer (Navios, Beckman Coulter) and based on the method described by Guttridge and colleagues . The vital dye 7‐aminoactinomycin‐D (7‐AAD) was used to identify nonviable cells and to calculate cell viability before and after processing.…”

Storage time affects umbilical cord blood viability

“…To support this, an additional 10 UCB samples cryopreserved within 24 hours of collection were tested using 7-AAD staining by flow cytometry. 17 The 10 donations had a collection volume of 149.80 ± 44.78 mL and their viability after cryopreservation was 47.03 ± 19.14% for granulocytes, 94.99 ± 4.54% for mononuclear cells, and 92.68 ± 8.83% for CD34+ cells. Far more important are the CD34+ stem and progenitor cells that make up less than 1% of the TNCs in UCB.…”

“…In our study, we observed a significant reduction in CD45+ cell viability both before and after cryopreservation, which is most likely attributed to the high granulocyte content. To support this, an additional 10 UCB samples cryopreserved within 24 hours of collection were tested using 7‐AAD staining by flow cytometry . The 10 donations had a collection volume of 149.80 ± 44.78 mL and their viability after cryopreservation was 47.03 ± 19.14% for granulocytes, 94.99 ± 4.54% for mononuclear cells, and 92.68 ± 8.83% for CD34+ cells.…”

“…UCB units were assayed both before and after cryopreservation to determine cell counts using a hematology analyzer (LH750, Beckman Coulter, High Wycombe, UK) and by flow cytometry based on the method described by Guttridge and coworkers . Viability of CD45+ cells and CD34+ cells was assessed by two methods.…”

“…UCB units were assayed both before and after cryopreservation to determine cell counts using a hematology analyzer (LH750, Beckman Coulter, High Wycombe, UK) and by flow cytometry based on the method described by Guttridge and coworkers. 17 Viability of CD45+ cells and CD34+ cells was assessed by two methods. First, all donations were tested by dye exclusion using 7-AAD in the flow cytometry assay used above 17 and, second, the proportion of apoptotic cells was assessed by flow cytometry using annexin V (Trevigen, Gaithersburg, MD) as described by the manufacturer.…”