PMID- 21673641
OWN - NLM
STAT- MEDLINE
DCOM- 20110830
LR  - 20211020
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 52
DP  - 2011 Jun 2
TI  - Generation and labeling of murine bone marrow-derived dendritic cells with Qdot 
      nanocrystals for tracking studies.
LID - 2785 [pii]
LID - 10.3791/2785 [doi]
AB  - Dendritic cells (DCs) are professional antigen presenting cells (APCs) found in 
      peripheral tissues and in immunological organs such as thymus, bone marrow, 
      spleen, lymph nodes and Peyer's patches. DCs present in peripheral tissues sample 
      the organism for the presence of antigens, which they take up, process and 
      present in their surface in the context of major histocompatibility molecules 
      (MHC). Then, antigen-loaded DCs migrate to immunological organs where they 
      present the processed antigen to T lymphocytes triggering specific immune 
      responses. One way to evaluate the migratory capabilities of DCs is to label them 
      with fluorescent dyes. Herewith we demonstrate the use of Qdot fluorescent 
      nanocrystals to label murine bone marrow-derived DC. The advantage of this 
      labeling is that Qdot nanocrystals possess stable and long lasting fluorescence 
      that make them ideal for detecting labeled cells in recovered tissues. To 
      accomplish this, first cells will be recovered from murine bone marrows and 
      cultured for 8 days in the presence of granulocyte macrophage-colony stimulating 
      factor in order to induce DC differentiation. These cells will be then labeled 
      with fluorescent Qdots by short in vitro incubation. Stained cells can be 
      visualized with a fluorescent microscopy. Cells can be injected into experimental 
      animals at this point or can be into mature cells upon in vitro incubation with 
      inflammatory stimuli. In our hands, DC maturation did not determine loss of 
      fluorescent signal nor does Qdot staining affect the biological properties of 
      DCs. Upon injection, these cells can be identified in immune organs by 
      fluorescent microscopy following typical dissection and fixation procedures.
FAU - Muccioli, Maria
AU  - Muccioli M
AD  - Molecular and Cell Biology Program, Ohio University, USA.
FAU - Pate, Michelle
AU  - Pate M
FAU - Omosebi, Omowaleola
AU  - Omosebi O
FAU - Benencia, Fabian
AU  - Benencia F
LA  - eng
GR  - R15 CA137499/CA/NCI NIH HHS/United States
GR  - R15 CA137499-01/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20110602
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - Bone Marrow Cells/*chemistry/cytology/*immunology
MH  - Dendritic Cells/*chemistry/cytology/immunology
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Microscopy, Fluorescence/*methods
MH  - *Quantum Dots
PMC - PMC3197051
EDAT- 2011/06/16 06:00
MHDA- 2011/08/31 06:00
PMCR- 2011/06/02
CRDT- 2011/06/16 06:00
PHST- 2011/06/16 06:00 [entrez]
PHST- 2011/06/16 06:00 [pubmed]
PHST- 2011/08/31 06:00 [medline]
PHST- 2011/06/02 00:00 [pmc-release]
AID - 2785 [pii]
AID - 10.3791/2785 [doi]
PST - epublish
SO  - J Vis Exp. 2011 Jun 2;(52):2785. doi: 10.3791/2785.