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. 2008 Jul 18;372(1):230-5.
doi: 10.1016/j.bbrc.2008.05.022. Epub 2008 May 13.

Upregulation of Runx2 and Osterix during in vitro chondrogenesis of human adipose-derived stromal cells

Jason T Rich  1 , Ivana Rosová, Jan A Nolta, Terence M Myckatyn, Linda J Sandell, Audrey McAlinden
Affiliations

Upregulation of Runx2 and Osterix during in vitro chondrogenesis of human adipose-derived stromal cells

Jason T Rich et al. Biochem Biophys Res Commun. .

Abstract

The aim of this study was to create a gene expression profile to better define the phenotype of human adipose-derived stromal cells (HADSCs) during in vitro chondrogenesis, osteogenesis and adipogenesis. A novel aspect of this work was the analysis of the same subset of genes during HADSC differentiation into all three lineages. Chondrogenic induction resulted in increased mRNA expression of Sox transcription factors, COL2A1,COL10A1, Runx2, and Osterix. This is the first report demonstrating significant upregulation in expression of osteogenesis-related transcription factors Runx2 and Osterix by TGF-beta3 induction of HADSCs during in vitro chondrogenesis. These findings suggest that the commonly-used chondrogenic induction reagents promote differentiation suggestive of hypertrophic chondrocytes and osteoblasts. We conclude that alternative strategies are required to induce efficient articular chondrocyte differentiation in order for HADSCs to be of clinical use in cartilage tissue engineering.

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Figures

Fig. 1
Fig. 1
Expression of adipogenic transcription factors PPAR-γ2 and C/EBP-α during differentiation of HADSCs. Checkered bars represent levels of mRNA from undifferentiated HADSCs (Day 0). Gray, white and black bars represent mRNA levels from HADSCs after induction of chondrogenesis, osteogenesis and adipogenesis, respectively. C=human primary articular chondrocytes, B=human cortical bone, A=human subcutaneous adipose tissue. * = P<0.05 relative to Day 0.
Fig. 2
Fig. 2
Expression of chondrogenesis related genes during differentiation of HADSCs. Checkered bars represent levels of mRNA from undifferentiated HADSCs (Day 0). Gray, white and black bars represent mRNA levels from HADSCs after induction of chondrogenesis, osteogenesis and adipogenesis, respectively. C=human primary articular chondrocytes, B=human cortical bone, A=human subcutaneous adipose tissue. * = P<0.05 relative to Day 0.
Fig. 3
Fig. 3
Expression of Indian Hedgehog (IHH) and type X collagen (COL10A1) during differentiation of HADSCs. Checkered bars represent levels of mRNA from undifferentiated HADSCs (Day 0). Gray, white and black bars represent mRNA levels from HADSCs after induction of chondrogenesis, osteogenesis and adipogenesis, respectively. C=human primary articular chondrocytes, B=human cortical bone, A=human subcutaneous adipose tissue. * = P<0.05 relative to Day 0.
Fig. 4
Fig. 4
Expression of osteogenesis related genes during HADSC differentiation. mRNA levels of transcription factors Runx2 and Osterix, and matrix markers alkaline phosphatase (Alk Phos) and type I collagen (COL1A1) were analyzed. Checkered bars represent levels of mRNA from undifferentiated HADSCs (Day 0). Gray, white and black bars represent mRNA levels from HADSCs after induction of chondrogenesis, osteogenesis and adipogenesis, respectively. C=human primary articular chondrocytes, B=human cortical bone, A=human subcutaneous adipose tissue. * = P<0.05 relative to Day 0.
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