PMID- 18320793
OWN - NLM
STAT- MEDLINE
DCOM- 20080508
LR  - 20141120
IS  - 2067-2993 (Print)
IS  - 2067-2993 (Linking)
VI  - 56
IP  - 4
DP  - 2007 Oct-Dec
TI  - [Experimental model of in vitro generation of human dendritic cells].
PG  - 183, 185-9
AB  - INTRODUCTION: Dendritic cells (DCs) are considered the most powerful 
      antigen-presenting cells (APC) playing a strategic role in triggering the immune 
      response. Monocytes are the physiological precursors of myeloid DCs. AIMS: This 
      paper aims to develop an in vitro experimental model of human dendritic cells 
      generation and phenotypic and functional analysis starting from CD14+ monocyte 
      precursors isolated from peripheral blood. MATERIALS AND METHODS: CD14+ monocytes 
      were isolated from venous peripheral blood which was harvested from consenting 
      healthy donors. By the means of GM-CSF and IL-4 immature DCs were generated from 
      monocytes. In the sixth day we added TNF-alpha and PGE-2 maturation factors and, 
      after 48 hours we harvested mature DCs. Their morphology was monitored by optical 
      microscopy, while the phenotypes were determined by flow cytometry. RESULTS: 
      After three days of culture, the monocytes developed a specific morphology, 
      gathering in clusters; cells presenting specific features start to detach in the 
      sixth day. Treatment with TNF-alpha and PGE-2 induces the maturation of DCs and 
      up-regulation of CD80, CD83 and CD86 expressions. Phenotypic analysis of mature 
      DCs was HLA-DR+++/CD86- CD80+++/CD14(-)/CD11a+/CD83+++. DCs obtained are able to 
      stimulate in vivo allolymphocyte capacity. Functional analysis test showed 87% 
      CD4+ CFSE+(DCs+PBMC) cells vs CD4+ CFSE+(PBMC) 15% cells. CONCLUSIONS: This 
      experiment developed an effective and viable way to generate DCs from PBMCs 
      exposed to GM-CSF and IL-4. The cells obtained presented morphological, 
      phenotypic and functional features as DCs described in the literature.
FAU - Oancea, Cristian
AU  - Oancea C
AD  - Catedra de Fiziologie Imunologie, Universitate de Medicina si Farmacie Victor 
      Babes Timisoara. oancea@umft.ro
FAU - Jiga, Janina
AU  - Jiga J
FAU - Tudorache, Voicu
AU  - Tudorache V
FAU - Paunescu, Virgil
AU  - Paunescu V
FAU - Mihalas, Georgeta
AU  - Mihalas G
FAU - Bunu, Carmen
AU  - Bunu C
FAU - Jiga, Lucian
AU  - Jiga L
LA  - rum
PT  - English Abstract
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
TT  - Obtinerea si cultivarea celulelor dendritice umane in vitro. Model experimental.
PL  - Romania
TA  - Pneumologia
JT  - Pneumologia (Bucharest, Romania)
JID - 100941067
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor)
SB  - IM
MH  - Adult
MH  - Cell Culture Techniques/methods
MH  - Cell Differentiation
MH  - Dendritic Cells/cytology/drug effects/immunology/*physiology
MH  - Female
MH  - Flow Cytometry
MH  - Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology
MH  - Humans
MH  - Immunophenotyping
MH  - Male
MH  - Monocytes/physiology
MH  - Phenotype
MH  - Tumor Necrosis Factor-alpha/pharmacology
EDAT- 2008/03/07 09:00
MHDA- 2008/05/09 09:00
CRDT- 2008/03/07 09:00
PHST- 2008/03/07 09:00 [pubmed]
PHST- 2008/05/09 09:00 [medline]
PHST- 2008/03/07 09:00 [entrez]
PST - ppublish
SO  - Pneumologia. 2007 Oct-Dec;56(4):183, 185-9.