Related Experiment Video
Updated: Mar 7, 2026
Isolation and Enrichment of Human Adipose-derived Stromal Cells for Enhanced Osteogenesis
Published on: January 12, 2015
RSPO3-LGR4 Regulates Osteogenic Differentiation Of Human Adipose-Derived Stem Cells Via ERK/FGF Signalling
Min Zhang1,2, Ping Zhang1,2, Yunsong Liu1,2
1Department of Prosthodontics, Peking University School and Hospital of Stomatology, Beijing, 100081, China.
Abstract:
The four R-spondins (RSPOs) and their three related receptors, LGR4, 5 and 6, have emerged as a major ligand-receptor system with critical roles in development and stem cell survival. However, the exact roles of the RSPO-LGR system in osteogenesis remain largely unknown. In the present study, we showed that RSPO3-shRNA increased the osteogenic potential of human adipose-derived stem cells (hASCs) significantly. Mechanistically, we demonstrated that RSPO3 is a negative regulator of ERK/FGF signalling. We confirmed that inhibition of the ERK1/2 signalling pathway blocked osteogenic differentiation in hASCs, and the increased osteogenic capacity observed after RSPO3 knockdown in hASCs was reversed by inhibition of ERK signalling. Further, silencing of LGR4 inhibited the activity of ERK signalling and osteogenic differentiation of hASCs. Most importantly, we found that loss of LGR4 abrogated RSPO3-regulated osteogenesis and RSPO3-induced ERK1/2 signalling inhibition. Collectively, our data show that ERK signalling works downstream of LGR4 and RSPO3 regulates osteoblastic differentiation of hASCs possibly via the LGR4-ERK signalling.
Insights
R-spondin 3 (RSPO3) negatively regulates osteogenesis in human stem cells by inhibiting ERK signaling. This pathway, involving LGR4, is crucial for RSPO3
Area of Science:
- Stem Cell Biology
- Molecular Biology
- Bone Biology
Background:
- The R-spondin (RSPO) and Leucine-rich repeat-containing G-protein coupled receptor (LGR) system is vital for development and stem cell maintenance.
- The specific role of the RSPO-LGR axis in osteogenesis, the process of bone formation, is not well understood.
Purpose of the Study:
- To investigate the role of R-spondin 3 (RSPO3) and its receptor LGR4 in the osteogenic differentiation of human adipose-derived stem cells (hASCs).
- To elucidate the underlying molecular mechanisms, particularly the involvement of the ERK/FGF signaling pathway.
Main Methods:
- Knockdown of RSPO3 using short hairpin RNA (shRNA) in hASCs.
- Inhibition of ERK1/2 signaling pathway using specific inhibitors.
- Silencing of LGR4 expression in hASCs.
- Assessment of osteogenic differentiation potential and ERK signaling activity.
Main Results:
- RSPO3 knockdown significantly enhanced the osteogenic potential of hASCs.
- RSPO3 acts as a negative regulator of ERK/FGF signaling.
- Inhibition of ERK1/2 signaling blocked osteogenic differentiation and reversed the effects of RSPO3 knockdown.
- LGR4 silencing inhibited ERK signaling and osteogenic differentiation.
- Loss of LGR4 abolished RSPO3-regulated osteogenesis and ERK1/2 signaling inhibition.
Conclusions:
- RSPO3 negatively regulates osteoblastic differentiation of hASCs.
- The LGR4-ERK signaling pathway is a key mediator of RSPO3's function in osteogenesis.
- ERK signaling operates downstream of LGR4 in the RSPO3-mediated regulation of osteogenesis.
More Related Videos
10:31Robust Differentiation of Human iPSCs into a Pure Population of Adipocytes to Study Adipocyte-Associated Disorders
Published on: February 9, 2022
08:28Expansion and Adipogenesis Induction of Adipocyte Progenitors from Perivascular Adipose Tissue Isolated by Magnetic Activated Cell Sorting
Published on: June 30, 2017