Semi‐automated flow cytometric analysis of CD34‐expressing hematopoietic cells in peripheral blood progenitor cell apheresis products

Abstract

The ProCOUNT kit and software for semi-automated data acquisition and analysis represents a further step toward standardization of CD34 cell quantitation in peripheral blood progenitor cell apheresis products. However, the occurrence of software warnings is high, and analysis or data reevaluation by experienced staff is still mandatory. Therefore, currently there is no definite advantage of the kit and software over the existing guidelines for CD34+ analysis in peripheral blood progenitor cell grafts.

Determining post‐thaw CD34+ cell dose of cryopreserved haematopoietic progenitor cells demonstrates high recovery and confirms their integrity

“…The kit consists of one test and one control absolute counting tube (TruCOUNT, BD Biosciences), each containing a known number of lyophilized fluorescent beads. In addition, ready‐to‐use reagents are provided [19]. These reagents, a nucleic acid dye in combination with a phycoerythrin (PE)‐labelled CD34+ monoclonal antibody (mAb; class III epitope, HPCA‐2, clone 8G12) and a CD45 peridinyl chlorophyll protein (PerCP) mAb (clone 2D1) for specific CD34 staining, and the nucleic acid dye in combination with a PE‐labelled immunoglobulin G1 (IgG1) control (mouse IgG1, clone X40) and the CD45 PerCP antibody for non‐specific staining (control reagent), are used in conjunction with the counting tubes.…”

Cord blood processing with an automated and functionally closed system

“…The enumeration of CD34+ cells was carried out with the ProCOUNT software (version 2.0, Becton Dickinson). The software's automated, specific multiple gating strategy was used, if no QC message was shown during the analysis 17 . Otherwise, the gates were reevaluated manually with ProCOUNT.…”

Preservation From Left Ventricular Remodeling by Front-Integrated Revascularization and Stem Cell Liberation in Evolving Acute Myocardial Infarction by Use of Granulocyte-Colony–Stimulating Factor (FIRSTLINE-AMI)

“…Stem cell concentration in peripheral blood was obtained according to the IGLD protocol 20 ; 100 L of peripheral blood (anticoagulant K3-EDTA) was incubated for 15 minutes on ice with both pretitered CD45-fluorescein-conjugated (clone 2D1) and CD34-phycoerythrin-conjugated (clone 8G12) monoclonal antibody of 10 L each. Similarly, control samples were incubated with fluorochromeconjugated isotypic control antibodies (clone X40), all purchased from Becton-Dickinson.…”