Clinical-grade manufacturing of blood dendritic cell subsets

The CliniMACS Prodigy® BloodDC PreDepletion System and the CliniMACS Prodigy BloodDC Enrichment and Culture System have been developed for the joint enrichment of dendritic cell (DC) subsets from non-mobilized apheresis products, followed by maturation and antigen loading. Steps of each system take place in the appropriate tubing set: CliniMACS Prodigy TS 320 for depletion and biotin labeling and CliniMACS Prodigy TS 510 for enrichment and cultivation. 

Enrichment and stimulation of primary blood DC subsets on the CliniMACS Prodigy platform

The CliniMACS Prodigy BloodDC PreDepletion System and the CliniMACS Prodigy BloodDC Enrichment and Culture System are designed to be used sequentially. First, the CliniMACS Prodigy BloodDC PreDepletion System performs the depletion of CD19+  and CD14+  cells and the labelling of CD1c+ cells with biotin. Subsequently, the CliniMACS Prodigy BloodDC Enrichment and Culture System allows the enrichment of the three blood DC subsets and, optionally, stimulation and antigen loading.
Purity and viability of blood DCs after enrichment with CliniMACS Prodigy Pre-Depletion System and CliniMACS Prodigy Enrichment and Culture System

Magnetic cell separation on the CliniMACS Prodigy is a simple method for efficiently isolating rare subsets of cells, such as blood DCs, in an automated and closed fashion. The CliniMACS Prodigy BloodDC PreDepletion System performs the depletion of CD19+ and CD14+ cells, and the labelling of CD1c+ cells with biotin. Subsequently, the CliniMACS Prodigy BloodDC Enrichment and Culture System allows the joint enrichment of the resultant blood DC subsets: CD304 (BDCA-4)+ plasmacytoid dendritic cells (pDCs), CD1c (BDCA-1)+ myeloid dendritic cells (cDC2s), and, optionally, CD141 (BDCA-3)+ myeloid dendritic cells (cDC1s).

Cytokine secretion and activation markers expression by blood DCs enriched with CliniMACS Prodigy Enrichment and Culture System

For fully functional antigen presentation capacity, DCs need to be stimulated with a specific danger signal, such as TLR-ligands, and loaded with antigens. 

After enrichment of DC subsets, the CliniMACS Prodigy BloodDC Enrichment and Culture System can perform a closed, controlled culture of DCs with the option of flexible addition of several stimulation reagents and antigen products, such as the PepTivator® Peptide Pools, at user-controlled intervals.

In-process and quality controls (IPC/QC) are required for consistent cell manufacturing. Integrated sampling pouches on the CliniMACS Tubing Sets allow for samples to be collected at any time during and at the end of the cell manufacturing process. Miltenyi Biotec provides complete solutions with its MACSQuant® Flow Cytometers, a time-saving and standardized staining procedure with StainExpress™ Dry Antibody Cocktails, and automated flow cytometry analysis with Express Modes for clinical flow cytometry.  

When using a MACSQuant Instrument, Express Modes allow for fully automated and standardized flow cytometry processes. The MACSQuant Instruments and wider MACS Flow Cytometry Portfolio are for research use only.

Application protocol and antibody panel
Immunophenotyping of dendritic cells from human whole blood using flow cytometry

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