Magnetic-Activated Cell Sorting: A Method to Isolate c-Kit Positive Cells
Published: April 30, 2023
Abstract
Source: Kesarwani, M., et al. Methods for Evaluating the Role of c-Fos and Dusp1 in Oncogene Dependence. J. Vis. Exp. (2019).
This video describes the technique of isolating c-kit positive cells from the bone marrow of a mouse. In the protocol, we will isolate c-kit positive cells from a transgenic mouse to validate c-Fos and Dusp1 alone or together as drug targets in leukemia.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. Isolation of c-Kit+ Cells from Bone Marrow Sacrifice the mice according to institutional guidelines. NOTE: Here, the mice were exposed to carbon dioxide (CO2) at an according to the AVMA recommended flow rate until death was determined by the cessation of respiration and heartbeat, fo…
Divulgaciones
The authors have nothing to disclose.
Materials
| Biological Materials | |||
| IMDM | Cellgro (corning) | 15-016-CVR | |
| Recombinant Murine SCF | Prospec | CYT-275 | |
| Recombinant Murine Flt3-Ligand | Prospec | CYT-340 | |
| Recombinant Murine IL-6 | Prospec | CYT-350 | |
| Recombinant Murine IL-7 | Peprotech | 217-17 | |
| FBS | Atlanta biological | S11150 | |
| PBS | Corning | 21040CV | |
| 70 μm nylon cell stariner | Becton Dickinson | 352350 | |
| LS Columns | Miltenyi | 130-042-401 | |
| EDTA | Ambion | AM9261 | |
| BSA | Sigma | A7906 | |
| Mortor pestle | Coor tek | 60316 and 60317 | |
| CD117 MicroBead Kit | Miltenyi | 130-091-224 | |
| Magnet Stand | Miltenyi |
Citar este artículo
Magnetic-Activated Cell Sorting: A Method to Isolate c-Kit Positive Cells. J. Vis. Exp. (Pending Publication), e20275, doi: (2023).