PMID- 19941120
OWN - NLM
STAT- MEDLINE
DCOM- 20100202
LR  - 20211028
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 595
DP  - 2010
TI  - Isolation of rat intestinal lymph DC.
PG  - 281-97
LID - 10.1007/978-1-60761-421-0_19 [doi]
AB  - Dendritic cells (DCs) migrate constitutively from the intestine via the lymph to 
      the mesenteric lymph nodes. These migrating intestinal lymph DCs (ilDCs) carry 
      antigens acquired in the intestine and play important roles in both the 
      initiation of immune responses and the maintenance of oral tolerance. The ilDC 
      population is made up of at least three functionally different DC subsets. Like 
      many DC populations, ilDCs are exquisitely sensitive to their environment, 
      changing their phenotype and maturing in response to the procedures associated 
      with their extraction from solid tissues. We have developed and refined a method 
      for collecting and purifying these DC subsets from rats, without inducing them to 
      mature. This method involves two separate surgical procedures, separated by at 
      least 6 weeks. Initially, mesenteric lymph nodes are removed. After the animals 
      have fully recovered we cannulate the thoracic duct and collect the ilDCs on ice, 
      minutes after they have left the lymph vessel. The DCs are then enriched using 
      magnetic beads and purified by flow cytometric sorting. We describe this method 
      here, including our recent refinements to limit the use of the restraining 
      "Bollman" cage.
FAU - Milling, Simon
AU  - Milling S
AD  - Glasgow Biomedical Research Centre, University of Glasgow, Glasgow, UK.
FAU - MacPherson, Gordon
AU  - MacPherson G
LA  - eng
GR  - BB/D012643/2/BB_/Biotechnology and Biological Sciences Research Council/United 
      Kingdom
GR  - G0900270/MRC_/Medical Research Council/United Kingdom
GR  - BB/D012643/1/BB_/Biotechnology and Biological Sciences Research Council/United 
      Kingdom
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
SB  - IM
MH  - Animals
MH  - Catheterization
MH  - Cells, Cultured
MH  - Dendritic Cells/*cytology
MH  - Flow Cytometry
MH  - Intestines/*cytology
MH  - Lymph Nodes/*cytology
MH  - Male
MH  - Models, Theoretical
MH  - Rats
MH  - Thoracic Duct
EDAT- 2009/11/27 06:00
MHDA- 2010/02/03 06:00
CRDT- 2009/11/27 06:00
PHST- 2009/11/27 06:00 [entrez]
PHST- 2009/11/27 06:00 [pubmed]
PHST- 2010/02/03 06:00 [medline]
AID - 10.1007/978-1-60761-421-0_19 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2010;595:281-97. doi: 10.1007/978-1-60761-421-0_19.