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…

Offenlegungen

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

Tags

Play Video

Diesen Artikel zitieren
In Vitro Mouse Bone Marrow Stromal Cell Differentiation into Adipocytes. J. Vis. Exp. (Pending Publication), e21527, doi: (2023).

View Video