Manufacturing of CD34  + HPC-enriched, high-purity mononuclear cell products from umbilical cord blood

AbstractThe purpose of this study was to explore methods of selectively enriching CD34  + haematopoietic progenitor cells (HPC) in mononuclear cell (MNC) preparations, and to outline a procedure for cryopreservation and thawing of manufactured material. Density gradient centrifugation of umbilical cord blood was achieved using Ficoll-Paque™ media at 1.077 g/mL and 1.065 g/mL d ensities and Leucosep preparation tubes. Post-process samples were analysed for CD34 + and MNC content. Finally, MNCs were frozen down at a concentration of 8.5 × 106 cells/mL in CryoStor CS10 using an Asymptote VIAFreeze controlled rate freezer at a rate of  − 2 °C per minute, then thawed and analysed for viability and recovery. Processing with 1.065 g/mL media selectively depleted non-HPC cell types, producing an approximately fourfold increase in CD34 + frequency (M ± 1SD = 1.4 ± 1.3%,P <  0.01) relative to the pre-process sample (M ± 1SD = 0.4 ± 0.3%), whereas 1.077 g/mL media produced only a twofold enrichment (0.7 ± 0.6,P <  0.01). This was not accompanied by any significant forfeit of CD34 + recovery (79 ± 32% vs. 78 ± 32% respectively;P = 0.87). The MNCs generated by the 1.065 g/mL procedure were of greater purity (96 ± 2%) than in the 1.077 g/mL procedure (80 ± 7%,P <  0.01). Post-thaw, MNC viability was 95 ± 1% and CD34 + viability was 98 ± 1%. Ultra-pure MNCs rich i...

Source: Cell and Tissue Banking - February 3, 2023 Category: Stem Cells Source Type: research