Related Experiment Video
Updated: May 31, 2026
Robust Differentiation of Human iPSCs into a Pure Population of Adipocytes to Study Adipocyte-Associated Disorders
Published on: February 9, 2022
Reactive oxygen species mediate adipocyte differentiation in mesenchymal stem cells
Yasunari Kanda1, Takashi Hinata, Sang Won Kang
1Department of Pharmacology, National Defense Medical College, Saitama 359-8513, Japan. kanda@nihs.go.jp
Aims:
Mesenchymal stem cells (MSC) have the potential to differentiate into various cell lineages, including adipocytes and osteoblasts. The formation of adipose tissue involves the commitment of MSC to the preadipocyte lineage and the differentiation of preadipocytes into mature adipocytes. In the present study, we investigated the involvement of reactive oxygen species (ROS) in adipocyte differentiation from MSC.
Main Methods:
ROS signaling was evaluated by the effects of antioxidant N-acetyl-l-cysteine (NAC) or shRNA against NAD(P)H oxidase in the multipotent mesenchymal stem cell line 10T1/2 cells. Intracellular ROS was measured using an H(2)DCF dye.
Key Findings:
We found that NAC blocked adipocyte differentiation in MSC. An H(2)DCF assay revealed that differentiation-inducing agents induced ROS generation. These data suggest that ROS is involved in adipocyte differentiation in MSC. Next, we examined the source of ROS. Knockdown of NAD(P)H oxidase 4 (Nox4) by RNA interference inhibited ROS production and adipocyte differentiation by differentiation-inducing agents. Furthermore, treatment with NAC blocked the transcriptional activation of CREB, and the expression of dominant-negative mutants of CREB inhibited adipocyte differentiation.
Significance:
The findings suggest that the increase in the intracellular ROS level via Nox4 mediates adipocyte differentiation through CREB in MSC. This data will provide new insight into the drug development for obesity.
