Bone Marrow-Derived Dendritic Cells Generation: A Method to Generate Dendritic Cells from Mouse Bone Marrow
Published: April 30, 2023
Abstract
Source:Nguyen, H. D., et al. Bone Marrow Transplantation Platform to Investigate the Role of Dendritic Cells in Graft-versus-Host Disease. J. Vis. Exp.. (2020).
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…
Divulgaciones
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) | GIBCO | 15140 | Media |
| RPMI 1640 | Thermo Fisher Scienctific | 11875-093 | Media |
Citar este artículo
Bone Marrow-Derived Dendritic Cells Generation: A Method to Generate Dendritic Cells from Mouse Bone Marrow. J. Vis. Exp. (Pending Publication), e20270, doi: (2023).