A method for the generation of large numbers of dendritic cells from CD34+ hematopoietic stem cells from cord blood.
Adjuvants, Immunologic
/ pharmacology
Antigens, CD34
/ metabolism
Bacterial Infections
/ immunology
Cell Count
Cell Differentiation
/ drug effects
Cells, Cultured
Cesarean Section
Culture Media
/ metabolism
Cytokines
/ metabolism
Dendritic Cells
Female
Fetal Blood
/ cytology
Granulocyte-Macrophage Colony-Stimulating Factor
/ metabolism
Hematopoietic Stem Cells
/ drug effects
Humans
Infant, Newborn
Lipopolysaccharides
/ immunology
Poly I-C
/ immunology
Primary Cell Culture
/ methods
Virus Diseases
/ immunology
CD34+ hematopoietic stem cells
Cord blood
Lipopolysaccharide
Maturation
Neonatal dendritic cells
Poly(I:C)
Journal
Journal of immunological methods
ISSN: 1872-7905
Titre abrégé: J Immunol Methods
Pays: Netherlands
ID NLM: 1305440
Informations de publication
Date de publication:
02 2020
02 2020
Historique:
received:
19
03
2019
revised:
06
11
2019
accepted:
07
11
2019
pubmed:
13
11
2019
medline:
30
9
2020
entrez:
13
11
2019
Statut:
ppublish
Résumé
Dendritic cells (DCs) play a central role in regulating innate and adaptive immune responses. It is well accepted that their regulatory functions change over the life course. In order to study DCs function during early life it is important to characterize the function of neonatal DCs. However, the availability of neonatal DCs is limited due to ethical reasons or relative small samples of cord blood making it difficult to perform large-scale experiments. Our aim was to establish a robust protocol for the generation of neonatal DCs from cord blood derived CD34+ hematopoietic stem cells. For the expansion of DC precursor cells we used a cytokine cocktail containing Flt-3 L, SCF, TPO, IL-3 and IL-6. The presence of IL-3 and IL-6 in the first 2 weeks of expansion culture was essential for the proliferation of DC precursor cells expressing CD14. After 4 weeks in culture, CD14+ precursor cells were selected and functional DCs were generated in the presence of GM-CSF and IL-4. Neonatal DCs were then stimulated with Poly(I:C) and LPS to mimic viral or bacterial infections, respectively. Poly(I:C) induced a higher expression of the maturation markers CD80, CD86 and CD40 compared to LPS. In line with literature data using cord blood DCs, our Poly(I:C) matured neonatal DCs cells showed a higher release of IL-12p70 compared to LPS matured neonatal DCs. Additionally, we demonstrated a higher release of IFN-γ, TNF-α, IL-1β and IL-6, but lower release of IL-10 in Poly(I:C) matured compared to LPS matured neonatal DCs derived from cord blood CD34+ hematopoietic stem cells. In summary, we established a robust protocol for the generation of large numbers of functional neonatal DCs. In line with previous studies, we showed that neonatal DCs generated form CD34+ cord blood progenitors have a higher inflammatory potential when exposed to viral than bacterial related stimuli.
Identifiants
pubmed: 31711888
pii: S0022-1759(19)30041-9
doi: 10.1016/j.jim.2019.112703
pmc: PMC6983936
pii:
doi:
Substances chimiques
Adjuvants, Immunologic
0
Antigens, CD34
0
Culture Media
0
Cytokines
0
Lipopolysaccharides
0
Granulocyte-Macrophage Colony-Stimulating Factor
83869-56-1
Poly I-C
O84C90HH2L
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
112703Subventions
Organisme : Medical Research Council
ID : G0800766
Pays : United Kingdom
Organisme : Medical Research Council
ID : G0900453
Pays : United Kingdom
Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.
Références
Transfusion. 2009 Oct;49(10):2109-21
pubmed: 19497054
Nat Rev Immunol. 2012 Jan 20;12(2):136-48
pubmed: 22266691
Proc Biol Sci. 2015 Dec 22;282(1821):20143085
pubmed: 26702035
J Exp Med. 2015 Mar 9;212(3):401-13
pubmed: 25687281
Allergy. 2011 May;66(5):579-87
pubmed: 21251015
Vaccine. 2016 Mar 18;34(13):1504-1514
pubmed: 26878294
Am J Pathol. 2011 Jul;179(1):248-58
pubmed: 21703407
Cancers (Basel). 2019 Feb 05;11(2):null
pubmed: 30764500
Front Physiol. 2018 Aug 02;9:877
pubmed: 30127748
J Allergy Clin Immunol. 2012 Apr;129(4):889-901
pubmed: 22464668
Blood. 1999 Apr 1;93(7):2244-52
pubmed: 10090933
Clin Exp Allergy. 2008 Mar;38(3):449-57
pubmed: 18177491
Expert Rev Clin Immunol. 2014 Sep;10(9):1171-84
pubmed: 25088080
Front Immunol. 2017 Jul 26;8:896
pubmed: 28798751
Eur J Immunol. 2009 Jan;39(1):26-35
pubmed: 19137537
Clin Sci (Lond). 2012 Sep;123(6):347-60
pubmed: 22264240
J Immunol. 2001 Feb 1;166(3):2141-6
pubmed: 11160266
Sci Rep. 2011;1:174
pubmed: 22355689
Front Immunol. 2017 May 01;8:499
pubmed: 28507549
J Immunol. 2002 Jun 15;168(12):6199-207
pubmed: 12055233
J Exp Med. 2001 Aug 20;194(4):519-27
pubmed: 11514607
J Leukoc Biol. 2002 Aug;72(2):321-9
pubmed: 12149423
J Immunol. 2017 Jan 15;198(2):557-563
pubmed: 28069750
Folia Biol (Praha). 2014;60(2):74-82
pubmed: 24785110