By Stephen P. Robinson, Andrew J. Stagg
Stephen P Robinson, MD, PhD, and Andrew Stagg, PhD, have introduced jointly quite a lot of time-proven equipment for learning those dendritic cells. lots of those without problems reproducible concepts care for the matter of acquiring enough dendritic cells for research, no matter if through isolation from a large vareity of tissues, or from numerous progenitor phone populations. different tools describe in step by step type the innovations regular for reading points of dendritic cells, starting from phone migration to antigen uptake and T cellphone stimulation. furthermore, a couple of strategies discover the sensible demanding situations interested by utilizing dendritic cells in a scientific environment to improve novel immunotherapeueutics.
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Additional info for Dendritic Cell Protocols (Methods in Molecular Medicine)
Remove the PBS-EDTA after the second wash. Add the VLDF cell suspension to the 5 mL polystyrene tube containing the prewashed magnetic beads and mix gently. Transfer the VLDF/magnetic bead suspension to a 1 mL round-bottom polypropylene CryoTube vial (Nunc; cat. no. , in a Dynal sampler mixer). Add 1 mL of PBS-EDTA-FCS to the VLDF/magnetic bead suspension, mix gently using a 1 mL micropipette, and transfer to a 5 mL polysterene tube (see Note 4). Place the tube in a Magnetic Particle Concentrator and leave it for 4 min.
Rat Spleen and Lymph DC 29 4 Isolation of Dendritic Cells from Rat Intestinal Lymph and Spleen Gordon MacPherson, Michelle Wykes, Fang-Ping Huang, and Chris Jenkins 1. 1. Lymph Dendritic Cells Dendritic cells (DC) are rare cells in peripheral tissues, and their isolation from tissues is fraught with problems. Thus, the proportion of DC within a tissue that is extracted is unknown, isolation procedures may select for subpopulations, and the isolation procedure itself may affect their properties.
Rat Spleen and Lymph DC 31 2. 1. Lymph Dendritic Cells 1. Specific pathogen-free rats around 6 wk of age are used for surgical lymphadenectomy (see Note 3). 2. Anesthetic apparatus: Any small animal apparatus suffices, but we use a halothane anaesthetic system (International Market Supply, Congleton, UK). No preoperative treatment is required. 3. Hair clippers (International Market Supply). 4. Surgical instruments including scalpel, scissors and dagger forceps (Holborn 4. , Margate, UK). 5. Sutures including absorbable 4/0 sutures (Ethicon) and needles (John Weiss, Milton Keynes, UK).