has knowledgeable contact person
generates dataset
uses tool
Protocols  >  Laboratory protocol  >  Monocyte enrichment by elut...

Monocyte enrichment by elutriation technology with ELUTRA(R)

Laboratory protocol

Leukapheresis products from cancer patients was used as a starting material for monocyte enrichment by elutriation technology with ELUTRA® (Gambro). The monocytes were cultured for five days to immature DC (imDC´s) and subsequently transfected with different constructs encoding for the oncogene Her-2/neu and as control PSA using the technology of Amaxa biosystems or an adeno virus Her-2 full length construct. Levels of Her-2 expression was highest following transfection with 3 different truncated Transmembrane-extracellular constructs, in particular a Her-2 rat-human chimeric one, while a full-length mutant Her-2 construct resulted in relatively low expression. Transfection with the adeno-Her-2 construct resulted in a very high expression level (80% positive cells) of Her-2 in combination with high cell viability (90%), which was in contrast to the relatively low expression observed with the same Her-2 construct as a pDNA construct transfected with the Amaxa based electroporation. We have concluded that when optimized the combination of leukapheresis and elutriation technologies, creates flexible platforms yielding large scale clinical grade enrichment of monocytes with high purity usefull for transfection with Her-2 or other cancer related gene products for DC based cell therapy and vaccination.

Protocol Steps:
  • Elutriation Leukapheresis products from cancer patients is used as a starting material for monocyte enrichment by elutriation technology with ELUTRA® (Gambro).
  • Monocyte culture The monocytes are cultured for five days to immature DC (imDC´s).
  • Transfection of immature DCs Transfection of immature DCs with different constructs encoding for the oncogene Her-2/neu and as control PSA using the technology of Amaxa biosystems or an adeno virus Her-2 full length construct.
Monocyte enrichment by elutriation technology with ELUTRA(R) Graph

created over 10 years ago (2 March 2009)    last modified over 7 years ago (28 September 2011)   [ RDF Rdf ]   [ RelFinder Relfinder ]