Fetal Mesenchymal Stromal Cells Differentiating towards Chondrocytes Acquire a Gene Expression Profile Resembling Human Growth Plate Cartilage

Abstract

We used human fetal bone marrow-derived mesenchymal stromal cells (hfMSCs) differentiating towards chondrocytes as an alternative model for the human growth plate (GP). Our aims were to study gene expression patterns associated with chondrogenic differentiation to assess whether chondrocytes derived from hfMSCs are a suitable model for studying the development and maturation of the GP. hfMSCs efficiently formed hyaline cartilage in a pellet culture in the presence of TGFβ3 and BMP6. Microarray and principal component analysis were applied to study gene expression profiles during chondrogenic differentiation. A set of 232 genes was found to correlate with in vitro cartilage formation. Several identified genes are known to be involved in cartilage formation and validate the robustness of the differentiating hfMSC model. KEGG pathway analysis using the 232 genes revealed 9 significant signaling pathways correlated with cartilage formation. To determine the progression of growth plate cartilage formation, we compared the gene expression profile of differentiating hfMSCs with previously established expression profiles of epiphyseal GP cartilage. As differentiation towards chondrocytes proceeds, hfMSCs gradually obtain a gene expression profile resembling epiphyseal GP cartilage. We visualized the differences in gene expression profiles as protein interaction clusters and identified many protein clusters that are activated during the early chondrogenic differentiation of hfMSCs showing the potential of this system to study GP development.

Endogenously Produced Indian Hedgehog Regulates TGFβ-Driven Chondrogenesis of Human Bone Marrow Stromal/Stem Cells

“…7A). Indeed, previous reports have shown an increase in IHH expression in response to TGFb stimulation [36,37] and SMAD4 has been shown to bind GTCTAGAC palindromic sequences within the 5¢ flanking region of the IHH gene, thus activating its transcription [38][39][40]. Concomitantly, SMAD2/3 interacts with SOX9 to increase its transcriptional activity, perhaps by increasing its interaction with CBP/p300 [41], and ERK1/2 is phosphorylated (data not shown), which has been shown to enhance the transcriptional activity of SMAD2/3 [42].…”

Molecular Validation of Chondrogenic Differentiation and Hypoxia Responsiveness of Platelet-Lysate Expanded Adipose Tissue–Derived Human Mesenchymal Stromal Cells

“…However, current protocols for chondrogenic differentiation efficiently create hypertrophic cartilage, which on implantation undergoes endochondral ossification and mineralization. 33 Several studies have attempted to understand the mechanisms by which MSCs can be programmed for chondrogenic differentiation while avoiding further progression into hypertrophic cartilage. 34,35 Mimicking the natural hypoxic environment of cartilage development under in vitro conditions is beneficial because it mediates metabolic programming of the chondrogenic fate of MSCs into different subtypes of hyaline cartilage.…”

Poly(γ-Glutamic Acid) as an Exogenous Promoter of Chondrogenic Differentiation of Human Mesenchymal Stem/Stromal Cells

“…36 Human neonatal bone marrow-derived MSCs were used due to their superior proliferation rate and differentiation capabilities resembling human growth plate cartilage, when compared to aged adult bone marrow-derived MSC. 77,78 As a source of differentiated cartilage cells, NCs were selected on account of their high degree of proliferation and stability of hyaline-cartilage phenotype, with increasing capacity for ECM production. 79 Although articular chondrocytes are the most studied chondrocyte source, NCs have the advantage of being able to be isolated under minimally invasive conditions with reduced site morbidity, 6,73,79 to be highly proliferative cells, with elevated and more reproducible chondrogenic capacity (both in vitro and ectopically in vivo), with increased capacity for ECM production (apparently donor ageindependent 79 and not affected by a joint pathological condition).…”