PMID- 29684429
OWN - NLM
STAT- MEDLINE
DCOM- 20190913
LR  - 20190913
IS  - 1872-7905 (Electronic)
IS  - 0022-1759 (Linking)
VI  - 458
DP  - 2018 Jul
TI  - Simultaneous in vitro generation of human CD34(+)-derived dendritic cells and 
      mast cells from non-mobilized peripheral blood mononuclear cells.
PG  - 63-73
LID - S0022-1759(17)30409-X [pii]
LID - 10.1016/j.jim.2018.04.005 [doi]
AB  - Dendritic cells (DCs) and mast cells (MCs) are key players of the immune system, 
      often coming in close proximity in peripheral tissues. The interplay of these 
      cells is, however, still poorly understood, especially with regards to human 
      cells. The reason for that is the absence of a well established in vitro human 
      cell-based study system that would allow a simultaneous preparation of both cell 
      types. In this study, we show a method for simultaneous generation of DCs and MCs 
      from CD34(+) stem cell progenitors that were isolated from the non-adherent 
      fraction of non-mobilized peripheral blood mononuclear cells of healthy donors. 
      We observed that combining stem cells factor (SCF), IL-3 and GM-CSF in serum-free 
      StemPro-34 medium allowed CD34(+) cells isolated from an equivalent of 450 ml of 
      peripheral blood to expand to 10-92 x 10(6) cells after 7 weeks of culturing. 
      These cultures comprised of 6-53% of DCs and 1-21% of MCs as determined by the 
      expression of, respectively, CD11c/HLA-DR or CD117/FcepsilonRI. The DCs were 
      CD1a(-)CD14(-), did not express co-stimulatory molecules CD80 and CD83 and 
      chemokine receptor CCR7. However, the DCs expressed co-stimulatory molecule CD86, 
      and had a capacity to uptake dextran, phagocyte latex particles and induce 
      alloreactivity. MCs, on the other hand, degranulated after crosslinking of 
      FcepsilonRI-bound IgE as determined by the externalization of CD107b. Collectively, our 
      data show that CD34(+)-derived human DCs and MCs can be generated in a single 
      culture using CD34(+) cells isolated from non-mobilized human peripheral blood 
      and that this method may allow ex vivo studies on DC-MC interplay in human 
      system.
CI  - Copyright (c) 2018 Elsevier B.V. All rights reserved.
FAU - Taborska, Pavla
AU  - Taborska P
AD  - Institute of Immunology, Charles University, 2nd Faculty of Medicine, University 
      Hospital Motol, Czech Republic.
FAU - Bartunkova, Jirina
AU  - Bartunkova J
AD  - Institute of Immunology, Charles University, 2nd Faculty of Medicine, University 
      Hospital Motol, Czech Republic.
FAU - Smrz, Daniel
AU  - Smrz D
AD  - Institute of Immunology, Charles University, 2nd Faculty of Medicine, University 
      Hospital Motol, Czech Republic. Electronic address: daniel.smrz@lfmotol.cuni.cz.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180421
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Antigens, CD34)
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Dextrans)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Stem Cell Factor)
RN  - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor)
SB  - IM
MH  - Antigens, CD34/*metabolism
MH  - Blood Buffy Coat/cytology
MH  - Cell Communication/immunology
MH  - Cell Degranulation/immunology
MH  - Cell Differentiation
MH  - Cell Separation/methods
MH  - Culture Media, Serum-Free/metabolism
MH  - Dendritic Cells/*immunology/metabolism
MH  - Dextrans/immunology
MH  - Flow Cytometry/methods
MH  - Granulocyte-Macrophage Colony-Stimulating Factor/metabolism
MH  - Healthy Volunteers
MH  - Humans
MH  - Leukocytes, Mononuclear
MH  - Mast Cells/*immunology
MH  - Peripheral Blood Stem Cells/*physiology
MH  - Primary Cell Culture/*methods
MH  - Recombinant Proteins/metabolism
MH  - Stem Cell Factor/metabolism
OTO - NOTNLM
OT  - CD34
OT  - Dendritic cells
OT  - Mast cells
EDAT- 2018/04/24 06:00
MHDA- 2019/09/14 06:00
CRDT- 2018/04/24 06:00
PHST- 2017/09/12 00:00 [received]
PHST- 2017/11/17 00:00 [revised]
PHST- 2018/04/18 00:00 [accepted]
PHST- 2018/04/24 06:00 [pubmed]
PHST- 2019/09/14 06:00 [medline]
PHST- 2018/04/24 06:00 [entrez]
AID - S0022-1759(17)30409-X [pii]
AID - 10.1016/j.jim.2018.04.005 [doi]
PST - ppublish
SO  - J Immunol Methods. 2018 Jul;458:63-73. doi: 10.1016/j.jim.2018.04.005. Epub 2018 
      Apr 21.