Optimization of leukocyte collection and monocyte isolation for dendritic cell culture

Abstract

Leukapheresis is the method of choice to collect monocytes for dendritic cell (DC) culture. Improvement of cell separators and cell collection software have enabled the collection of 10(9) monocytes for the generation of monocyte-derived DCs, which is sufficient to prepare a DC vaccine series. However, leukapheresis works with the technique of differential centrifugation which is not applicable to selectively collect mononuclear cells of similar density. After leukapheresis, thus, additional preparation steps are required to isolate and enrich the desired monocyte population. The cell isolation and cultivation techniques depend on the quality of the original leukocyte harvest due to the monocyte yield and the content of residual erythrocytes and platelets. Monocyte elutriation from the leukapheresis product shows a high monocyte recovery of 80%. However, only 30% of the isolated monocytes can be developed into mature DCs. The factors responsible for DC maturation and the development of different DC subsets are the subject of current research.