R. Yerushalmi, I. McNiece, C. Childs, E. J. Shpall
ISHAGE San Diego 2000 Abstrct - abstract # 106
Dendritic cell expansion
We compared DC expansion in serum-containing to serum-free media. CD34+ PBPCs were cultured in Teflon bags using different medias: a-MEM + 20% FBS, Amgen Defined Media, and QBSF-60, supplemented with 25ng/ml rh-GM-CSF, 100ng/ml rh-SCF and 50ng/ml rh- TNFa. The cells were fed every 7 days with fresh media and double the concentration of cytokines. Cells were harvested after 7, 10, 14, and 17 days, and were examined for DC morpholoqv by light microscopy and for DC phenotype by flow cytometry.
The a-MEM with 20% FBS appears to generate a higher number of DCs (9e6) when compared to Amgen Defined Media (7.4e6), or QBSf- 60 (l.8e6 ). FBS may be necessary for maximal generation of DC from CD34+ PBPC. Further studies are in progress to confirm these data. Published clinical studies report 5-20e6 DCs infused per treatment. Using a-MEM with 20% FBS in the culture conditions described, substantially larger DC numbers were generated. A typical CD34+ PBPC product can generate approximately 3.6eS DCs. Clinical studies to define the clinical efficacy of large scale DC expansion using these conditions are planned.