In Vitro Mouse Bone Marrow Stromal Cell Differentiation into Adipocytes

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…

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