Source: Maridas, D. E. et al., Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice. J. Vis. Exp. (2018)
This video demonstrates the differentiation of bone marrow stromal cells (BMSCs) into adipocytes. In the two-step process, in the presence of specific growth factors and hormones, undifferentiated BMSCs first differentiate into preadipocytes. The second step involving treatment with glucose and insulin leads to the formation of mature adipocytes with multiple lipid droplets.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Collection Tubes Preparation
2. Harvest of Bones from Mice
3. Cell Suspension
4. Plating and Culture of Mixed Populations of Bone Marrow Cells (Total BMCs)
NOTE: Cells are cultured at 37 °C in an incubator with 5% CO2.
5. Plating and Culture of the Split Population of BMSCs (Adherent BMSCs)
NOTE: Cells are cultured at 37 °C in an incubator with 5% CO2.
6. Differentiation of BMSCs
The authors have nothing to disclose.
200μL pipet tips | Rainin | 17014401 | |
1.5mL centrifuge tubes | USA Scientific | 1615-5500 | |
15mL conical tube | VWR | 89039-668 | |
50mL conical tube | VWR | 89039-660 | |
Phosphate-Buffered Saline (PBS) | Sigma-Aldrich | 21-040-CM | |
Ethanol | Fisher Science | 04-355-451 | |
Dissection tools | |||
70μm filters | BD Falcon | 352350 | |
0.25% trypsin | Gibco | 25200 | |
Kimwipe | VWR | 82003-820 | |
MEMα | Gibco | 12571 | |
Fetal Bovine Serum | VWR | 97068-085 | |
Penicillin/Streptomycin | Invitrogen | 15140-122 | |
DMEM High Glucose | Sigma-Aldrich | D5796 | |
Rosiglitazone | Cayman Chemical | 717410 | |
Insulin | Sigma-Aldrich | I6634 | |
IBMX | Sigma-Aldrich | I5879 | |
Dexamethasone | Sigma-Aldrich | D4902 | |
Axio Observer inverter microscope | Zeiss |