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Updated: May 9, 2026
Chondrogenic Differentiation Induction of Adipose-derived Stem Cells by Centrifugal Gravity
Published on: February 24, 2017
Chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells treated by GSK-3 inhibitors
Mohamadreza Baghaban Eslaminejad1, Negar Karimi, Maryam Shahhoseini
1Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 16635-148, Tehran, Iran, eslami@royaninstitute.org.
Abstract:
A study of the cartilage differentiation of mesenchymal stem cells (MSCs) would be of particular interest since one strategy for cell-based treatment of cartilage defects emphasizes the use of cells that are in a differentiated state. The present study has attempted to evaluate the effects of two well-known glycogen synthase kinase-3 inhibitors, including lithium chloride (LiCl) and SB216763 on a human marrow-derived MSC (hMSC) chondrogenic culture. Passaged-3 MSCs were condensed into small pellets and cultivated in the following groups based on the supplementation of chondrogenic medium: transforming growth factor (TGF)-β1, TGF-β1 + LiCl, TGF-β1 + SB216763, TGF-β3, TGF-β3 + LiCl, and TGF-β3 + SB216763. The cultures were maintained for 21 days and then analyzed for expression of Sox9, aggrecan, collagen II, β-catenin, and axin genes. Deposition of glycosaminoglycan (GAG) in the cartilage matrix was also measured for certain cultures. The presence of both LiCl and SB216763 along with TGF-β in the MSC chondrogenic culture led to the up-regulation of cartilage-specific genes. TGF-β3 appeared much better than TGF-β1. Based on our findings, SB216763 was more effective in up-regulation of cartilage-specific genes. These chondrogenic effects appeared to be mediated through the Wnt signaling pathway since β-catenin and axin tended to be up-regulated and down-regulated, respectively. In the culture with SB216763 + TGF-β3, significantly more GAG was deposited (P < 0.05). In conclusion, addition of either SB216763 or LiCl to hMSC chondrogenic culture up-regulates cartilage-specific gene expression and enhances GAG deposition in the culture.
Insights
Two glycogen synthase kinase-3 inhibitors, lithium chloride (LiCl) and SB216763, enhance human marrow-derived mesenchymal stem cell (hMSC) chondrogenesis. These compounds up-regulate cartilage-specific gene expression and glycosaminoglycan deposition, aiding cartilage repair strategies.
Area of Science:
- Biomedical Engineering
- Stem Cell Biology
- Regenerative Medicine
Background:
- Cell-based therapies for cartilage defects often require differentiated cells.
- Mesenchymal stem cells (MSCs) are a promising source for cartilage regeneration.
- Understanding factors that promote MSC chondrogenesis is crucial for therapeutic development.
Purpose of the Study:
- To evaluate the effects of glycogen synthase kinase-3 (GSK-3) inhibitors, lithium chloride (LiCl) and SB216763, on human marrow-derived MSC (hMSC) chondrogenic culture.
- To compare the efficacy of transforming growth factor-beta 1 (TGF-β1) and TGF-β3 in combination with GSK-3 inhibitors.
- To investigate the role of the Wnt signaling pathway in mediating chondrogenic effects.
Main Methods:
- Human marrow-derived MSCs were cultured as pellets in chondrogenic medium supplemented with TGF-β1 or TGF-β3, with or without LiCl or SB216763.
- Cultures were maintained for 21 days.
- Analysis included gene expression (Sox9, aggrecan, collagen II, β-catenin, axin) and glycosaminoglycan (GAG) deposition.
Main Results:
- Both LiCl and SB216763, when combined with TGF-β, significantly upregulated cartilage-specific genes.
- TGF-β3 was more effective than TGF-β1.
- SB216763 demonstrated superior upregulation of cartilage-specific genes and significantly enhanced GAG deposition in combination with TGF-β3.
- Wnt signaling pathway activation was suggested by β-catenin upregulation and axin downregulation.
Conclusions:
- Addition of SB216763 or LiCl to hMSC chondrogenic cultures enhances cartilage-specific gene expression.
- These GSK-3 inhibitors promote GAG deposition, a key component of cartilage matrix.
- SB216763, particularly with TGF-β3, shows significant potential for improving MSC-based cartilage regeneration therapies.
