PMID- 14594514
OWN - NLM
STAT- MEDLINE
DCOM- 20040609
LR  - 20171116
IS  - 1525-8165 (Print)
IS  - 1525-8165 (Linking)
VI  - 12
IP  - 5
DP  - 2003 Oct
TI  - Dendritic cell culture: a simple closed culture system using ficoll, monocytes, 
      and a table-top centrifuge.
PG  - 575-85
AB  - Dendritic cells (DCs) are potent antigen-presenting cells involved in the 
      induction of T cell-mediated immune responses and as such have emerged as 
      important candidates for cellular-based therapies. Critical to safe clinical use 
      is the easy manipulation of DCs and their precursors in a closed system. We have 
      developed a serum-free, closed culture system applying a simple wash-Ficoll 
      centrifugation method to reduce platelet and red blood cell (RBC) contamination. 
      This procedure optimized adherence of monocytes (44 +/- 10.9% recovery, >85% 
      expressed CD14(+)/CD163(+)) for the generation of DCs from mononuclear cell (MNC) 
      apheresis units. Most RBCs and up to 98% of platelets were removed. Following 
      density sedimentation, cell viability remained high (98 +/- 2%) with only minimal 
      loss of monocytes (3 +/- 3%). Importantly, Ficoll-treated monocytes retained 
      their ability to differentiate to mature DCs demonstrated by morphology, 
      phenotype (MHC class II(+), CD1a(+), CD80(+), CD86(+), and CD83(+)), ability to 
      stimulate mixed lymphocyte responses (MLR), present antigen, and produce 
      interleukin-12 (IL-12). Nonadherent CD3(+) (80 +/- 4%) were also isolated for 
      functional assays. Ficoll can be easily incorporated into a simple 
      adherence-based closed system for collection of lymphocytes and adherent 
      monocytes for DC culture. The procedure is relatively fast (effective working 
      time 5-6 h), does not impair monocyte function or induce substantial cell 
      activation, and can be performed economically using equipment found in a typical 
      blood banking environment.
FAU - Celluzzi, Christina M
AU  - Celluzzi CM
AD  - Blood and Cell Therapy Development Department, Jerome H. Holland Laboratory for 
      the Biomedical Sciences, American Red Cross, Rockville, MD 20855-2743, USA. 
      Celluzzi@usa.redcross.org
FAU - Welbon, Craig
AU  - Welbon C
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Hematother Stem Cell Res
JT  - Journal of hematotherapy & stem cell research
JID - 100892915
RN  - 0 (Annexins)
RN  - 0 (Antigens, CD)
RN  - 0 (Antigens, Surface)
RN  - 0 (Cytokines)
RN  - 0 (Interleukins)
RN  - 0 (Lipopolysaccharide Receptors)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Phytohemagglutinins)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 207137-56-2 (Interleukin-4)
RN  - 25702-74-3 (Ficoll)
RN  - 82115-62-6 (Interferon-gamma)
RN  - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor)
SB  - IM
MH  - Annexins/analysis
MH  - Antigen Presentation/physiology
MH  - Antigens, CD/analysis
MH  - Antigens, Surface/analysis
MH  - Apoptosis
MH  - Blood Component Removal
MH  - Cell Adhesion
MH  - Cell Count
MH  - Cell Culture Techniques/instrumentation/*methods
MH  - Cell Differentiation/*physiology
MH  - Cell Survival
MH  - Centrifugation
MH  - Cytokines/analysis
MH  - Dendritic Cells/cytology/*metabolism
MH  - Ficoll/*chemistry/pharmacology
MH  - Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology
MH  - Humans
MH  - Interferon-gamma/analysis/pharmacology
MH  - Interleukin-4/pharmacology
MH  - Interleukins/analysis
MH  - Leukocytes, Mononuclear/cytology/metabolism
MH  - Lipopolysaccharide Receptors/analysis
MH  - Lipopolysaccharides/pharmacology
MH  - Lymphocyte Activation/physiology
MH  - Monocytes/cytology/drug effects/*metabolism
MH  - Phytohemagglutinins/pharmacology
MH  - T-Lymphocytes/cytology/drug effects/metabolism
MH  - Tumor Necrosis Factor-alpha/analysis/pharmacology
EDAT- 2003/11/05 05:00
MHDA- 2004/06/21 10:00
CRDT- 2003/11/05 05:00
PHST- 2003/11/05 05:00 [pubmed]
PHST- 2004/06/21 10:00 [medline]
PHST- 2003/11/05 05:00 [entrez]
AID - 10.1089/152581603322448286 [doi]
PST - ppublish
SO  - J Hematother Stem Cell Res. 2003 Oct;12(5):575-85. doi: 
      10.1089/152581603322448286.