This video describes a protocol for the generation of dendritic cells–key antigen-presenting cells that activate T cells–from the bone marrow of an adult mouse.
Protocol
1. Generate bone marrow-derived dendritic cells (BM-DCs). Isolate bone marrow from the femurs and tibias of WT or factor B (fB)-/- mice on the B6 background. Briefly, collect the tibia and femur from both legs using forceps and scissors. Put them in ice-cold RPMI containing 1% FCS and 100 U/mL penicillin/streptomycin and 2 mM L-glutamine (1% RPMI) in 50 mL conical tubes. Clean the femur and the tibia bones thoroughly by removing all the muscle tissues using forceps and scissors. Transfer the f…
Declarações
The authors have nothing to disclose.
Materials
10X PBS
Fisher Scientific
BP3994
MACS buffer
B6 fB-/- mice
University of Central Florida
In house
Recipients
B6.Ly5.1 (CD45.1 + ) mice
Charles River
546
Donors
BALB/c mice
Charles River
28
Transplant recipients
C57BL/6 mice
Charles River
27
Donors/Recipients
Fetal Bovine Serum (FBS)
Atlanta Bilogicals R&D system
D17051
Cell Culture
Lipopolysacharide (LPS)
Millipore Sigma
L4391-1MG
DC mature
New Brunswick Galaxy 170R incubator
Eppendorf
Galaxy 170 R
Cell Culture
Penicilin+streptomycinPenicillin/ Streptomycin (10,000 units penicillin / 10,000 mg/ml strep)
Bone Marrow-Derived Dendritic Cells Generation: A Method to Generate Dendritic Cells from Mouse Bone Marrow. J. Vis. Exp. (Pending Publication), e20270, doi: (2023).