PMID- 27142009
OWN - NLM
STAT- MEDLINE
DCOM- 20171213
LR  - 20180110
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1423
DP  - 2016
TI  - The Isolation and Enrichment of Large Numbers of Highly Purified Mouse Spleen 
      Dendritic Cell Populations and Their In Vitro Equivalents.
PG  - 61-87
LID - 10.1007/978-1-4939-3606-9_5 [doi]
AB  - Dendritic cells (DCs) form a complex network of cells that initiate and 
      orchestrate immune responses against a vast array of pathogenic challenges. 
      Developmentally and functionally distinct DC subtypes differentially regulate 
      T-cell function. Importantly it is the ability of DC to capture and process 
      antigen, whether from pathogens, vaccines, or self-components, and present it to 
      naive T cells that is the key to their ability to initiate an immune response. 
      Our typical isolation procedure for DC from murine spleen was designed to 
      efficiently extract all DC subtypes, without bias and without alteration to their 
      in vivo phenotype, and involves a short collagenase digestion of the tissue, 
      followed by selection for cells of light density and finally negative selection 
      for DC. The isolation procedure can accommodate DC numbers that have been 
      artificially increased via administration of fms-like tyrosine kinase 3 ligand 
      (Flt3L), either directly through a series of subcutaneous injections or by 
      seeding with an Flt3L secreting murine melanoma. Flt3L may also be added to bone 
      marrow cultures to produce large numbers of in vitro equivalents of the spleen DC 
      subsets. Total DC, or their subsets, may be further purified using 
      immunofluorescent labeling and flow cytometric cell sorting. Cell sorting may be 
      completely bypassed by separating DC subsets using a combination of fluorescent 
      antibody labeling and anti-fluorochrome magnetic beads. Our procedure enables 
      efficient separation of the distinct DC subsets, even in cases where mouse 
      numbers or flow cytometric cell sorting time is limiting.
FAU - Vremec, David
AU  - Vremec D
AD  - The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, 
      Parkville, VIC, 3050, Australia. vremec@wehi.edu.au.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Membrane Proteins)
RN  - 0 (flt3 ligand protein)
SB  - IM
MH  - Animals
MH  - Bone Marrow Cells/cytology
MH  - Cell Separation/methods
MH  - Cells, Cultured
MH  - Dendritic Cells/*cytology/drug effects/immunology
MH  - Flow Cytometry/methods
MH  - Immunomagnetic Separation/*methods
MH  - Membrane Proteins/administration & dosage/pharmacology
MH  - Mice
MH  - Spleen/*cytology/drug effects
OTO - NOTNLM
OT  - Conventional DCs
OT  - DC expansion
OT  - DC purification
OT  - DC subtypes
OT  - Dendritic cell
OT  - Plasmacytoid DCs
EDAT- 2016/05/05 06:00
MHDA- 2017/12/14 06:00
CRDT- 2016/05/05 06:00
PHST- 2016/05/05 06:00 [entrez]
PHST- 2016/05/05 06:00 [pubmed]
PHST- 2017/12/14 06:00 [medline]
AID - 10.1007/978-1-4939-3606-9_5 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2016;1423:61-87. doi: 10.1007/978-1-4939-3606-9_5.