In Vitro Mouse Bone Marrow Stromal Cell Differentiation into Adipocytes
Published: July 31, 2023
Abstract
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.
Protocol
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 Make the BMSC culture media by supplementing Minimum Essential Medium α (MEM α) with 10% Fetal Bovine Serum and 1% of Penicillin/Streptomycin. Place 100 µL of BMSC culture media in 1.5 mL microcentrifuge tubes. 3 bones will usually fit in one single tube so prepare a…
開示
The authors have nothing to disclose.
Materials
| 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 |
