Isolation and Chondrogenic Differentiation of Porcine Perichondrial Progenitor Cells for the Purpose of Cartilage Tissue Engineering

Derks, Mareike; Sturm, Theresa; Haverich, Axel; Hilfiker, Andres

doi:10.1159/000354897

Cited by 24 publications

(18 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cell masses were then treated with no EPO, 40 ng/ml EPO or EPO block peptide (500 ng/ml, Santa Cruz) in chondrogenic medium supplemented with dexamethasone (100 nM, Sigma-Aldrich), ascorbic acid (50 µg/ml, Sigma-Aldrich), sodium pyruvate (100 µg/ml, Gibco), ITS+Premix (50 mg/ml, BD) for 7 days following previous procedures [48].…”

Section: Methodsmentioning

confidence: 99%

EPO Promotes Bone Repair through Enhanced Cartilaginous Callus Formation and Angiogenesis

Lin

Zhang

et al. 2014

PLoS ONE

View full text Add to dashboard Cite

Erythropoietin (EPO)/erythropoietin receptor (EPOR) signaling is involved in the development and regeneration of several non-hematopoietic tissues including the skeleton. EPO is identified as a downstream target of the hypoxia inducible factor-α (HIF-α) pathway. It is shown that EPO exerts a positive role in bone repair, however, the underlying cellular and molecular mechanisms remain unclear. In the present study we show that EPO and EPOR are expressed in the proliferating, pre-hypertrophic and hypertrophic zone of the developing mouse growth plates as well as in the cartilaginous callus of the healing bone. The proliferation rate of chondrocytes is increased under EPO treatment, while this effect is decreased following siRNA mediated knockdown of EPOR in chondrocytes. EPO treatment increases biosynthesis of proteoglycan, accompanied by up-regulation of chondrogenic marker genes including SOX9, SOX5, SOX6, collagen type 2, and aggrecan. The effects are inhibited by knockdown of EPOR. Blockage of the endogenous EPO in chondrocytes also impaired the chondrogenic differentiation. In addition, EPO promotes metatarsal endothelial sprouting in vitro. This coincides with the in vivo data that local delivery of EPO increases vascularity at the mid-stage of bone healing (day 14). In a mouse femoral fracture model, EPO promotes cartilaginous callus formation at days 7 and 14, and enhances bone healing at day 28 indexed by improved X-ray score and micro-CT analysis of microstructure of new bone regenerates, which results in improved biomechanical properties. Our results indicate that EPO enhances chondrogenic and angiogenic responses during bone repair. EPO's function on chondrocyte proliferation and differentiation is at least partially mediated by its receptor EPOR. EPO may serve as a therapeutic agent to facilitate skeletal regeneration.

show abstract

Section: Methodsmentioning

confidence: 99%

EPO Promotes Bone Repair through Enhanced Cartilaginous Callus Formation and Angiogenesis

Lin

Zhang

et al. 2014

PLoS ONE

View full text Add to dashboard Cite

show abstract

“…Tissue-derived stem/progenitor cells exhibit stem-cell-like qualities, such as self-renewal and multipotency, yet are embedded within the target tissue in niches and are primed to differentiate to that tissue (Jayasuriya and Chen, 2015). The identification of resident progenitor cell populations in articular cartilage (Dowthwaite et al, 2004;Williams et al, 2010) as well as in auricular and tracheal perichondrium (Derks et al, 2013;Kobayashi et al, 2011b;Togo et al, 2006) has opened up new pathways for cartilage tissue engineering. Like MSCs, which can undergo up to 70 population doublings (Christodoulou et al, 2013), cartilage progenitor/stem cells retain proliferative ability for up to 60 population doublings (Williams et al, 2010), demonstrating potential for accumulating large cell numbers starting from a single cell.…”

Section: Introductionmentioning

confidence: 99%

Progenitor cells in auricular cartilage demonstrate cartilage-forming capacity in 3D hydrogel culture

Otto¹,

Levato²,

Webb³

et al. 2018

eCM

View full text Add to dashboard Cite

Paramount for the generation of auricular structures of clinically-relevant size is the acquisition of a large number of cells maintaining an elastic cartilage phenotype, which is the key in producing a tissue capable of withstanding forces subjected to the auricle. Current regenerative medicine strategies utilize chondrocytes from various locations or mesenchymal stromal cells (MSCs). However, the quality of neo-tissues resulting from these cell types is inadequate due to inefficient chondrogenic differentiation and endochondral ossification, respectively. Recently, a subpopulation of stem/progenitor cells has been identified within the auricular cartilage tissue, with similarities to MSCs in terms of proliferative capacity and cell surface biomarkers, but their potential for tissue engineering has not yet been explored. This study compared the in vitro cartilage-forming ability of equine auricular cartilage progenitor cells (AuCPCs), bone marrow-derived MSCs and auricular chondrocytes in gelatin methacryloyl (gelMA)-based hydrogels over a period of 56 d, by assessing their ability to undergo chondrogenic differentiation. Neocartilage formation was assessed through gene expression profiling, compression testing, biochemical composition and histology. Similar to MSCs and chondrocytes, AuCPCs displayed a marked ability to generate cartilaginous matrix, although, under the applied culture conditions, MSCs outperformed both cartilage-derived cell types in terms of matrix production and mechanical properties. AuCPCs demonstrated upregulated mRNA expression of elastin, low expression of collagen type X and similar levels of proteoglycan production and mechanical properties as compared to chondrocytes. These results underscored the AuCPCs' tissue-specific differentiation potential, making them an interesting cell source for the next generation of elastic cartilage tissue-engineered constructs.

show abstract

“…In addition, the RT-PCR analyses showed an increase in the gene expression of specific marker genes for mature cartilage, such as SOX-9 and collagen II over 12 weeks in vivo (Figure 9). However, it cannot be excluded that progenitor cells from the perichondrium of the remaining local cartilage are responsible for the generation of neo-cartilage within the PUfibrin construct since there is strong evidence for the existence of multipotent progenitor cells in the perichondrium [57][58][59][60][61]. The ASCs were not labelled before implantation; thus, it is not possible to distinguish between implanted ASCs and resident cells in the present study.…”

Section: Discussionmentioning

confidence: 80%

Differentiation Behaviour of Adipose-Derived Stromal Cells (ASCs) Seeded on Polyurethane-Fibrin Scaffolds In Vitro and In Vivo

et al. 2021

View full text Add to dashboard Cite

Adipose-derived stromal cells (ASCs) are a promising cell source for tissue engineering and regenerative medicine approaches for cartilage replacement. For chondrogenic differentiation, human (h)ASCs were seeded on three-dimensional polyurethane (PU) fibrin composites and induced with a chondrogenic differentiation medium containing TGF-ß3, BMP-6, and IGF-1 in various combinations. In addition, in vitro predifferentiated cell-seeded constructs were implanted into auricular cartilage defects of New Zealand White Rabbits for 4 and 12 weeks. Histological, immunohistochemical, and RT-PCR analyses were performed on the constructs maintained in vitro to determine extracellular matrix (ECM) deposition and expression of specific cartilage markers. Chondrogenic differentiated constructs showed a uniform distribution of cells and ECM proteins. RT-PCR showed increased gene expression of collagen II, collagen X, and aggrecan and nearly stable expression of SOX-9 and collagen I. Rabbit (r)ASC-seeded PU-fibrin composites implanted in ear cartilage defects of New Zealand White Rabbits showed deposition of ECM with structures resembling cartilage lacunae by Alcian blue staining. However, extracellular calcium deposition became detectable over the course of 12 weeks. RT-PCR showed evidence of endochondral ossification during the time course with the expression of specific marker genes (collagen X and RUNX-2). In conclusion, hASCs show chondrogenic differentiation capacity in vitro with the expression of specific marker genes and deposition of cartilage-specific ECM proteins. After implantation of predifferentiated rASC-seeded PU-fibrin scaffolds into a cartilage defect, the constructs undergo the route of endochondral ossification.

show abstract

Isolation and Chondrogenic Differentiation of Porcine Perichondrial Progenitor Cells for the Purpose of Cartilage Tissue Engineering

Cited by 24 publications

References 41 publications

EPO Promotes Bone Repair through Enhanced Cartilaginous Callus Formation and Angiogenesis

EPO Promotes Bone Repair through Enhanced Cartilaginous Callus Formation and Angiogenesis

Progenitor cells in auricular cartilage demonstrate cartilage-forming capacity in 3D hydrogel culture

Differentiation Behaviour of Adipose-Derived Stromal Cells (ASCs) Seeded on Polyurethane-Fibrin Scaffolds In Vitro and In Vivo

Contact Info

Product

Resources

About