Modulation of Chondrocytic Properties of Fat-Derived Mesenchymal Cells in Co-Cultures with Nucleus Pulposus

Abstract

Human subcutaneous fat-derived mesenchymal cells recently have been shown to have the potential to differentiate in vitro into a variety of cell types, including adipocytes, osteoblasts, chondrocytes, and myoblasts. This effect suggests that fat tissue may serve as an abundant and easily acquired source of multipotent cells for tissue engineering. The multipotential characteristics of fat-derived mesenchymal cells from the inguinial fat pad of rabbit have not been clearly defined. In this study we have isolated a population of mesenchymal cells from inguinal fat from adult New Zealand white rabbits. The cells that were maintained under various differentiation conditions were shown to differentiate in vitro into adipocytes, osteoblasts, or chondrocytes; this differentiation was demonstrated using gene expression for tissue-specific proteins. We also co-cultured the cells with intervertebral disk tissue from the nucleus pulpous or from the annulus fibrosus. The fat-derived cells co-cultured with nucleus pulposus showed an increase in expression of type II collagen and aggrecan genes, compared with cells in alginate alone and cells co-cultured with annulus fibrosus. The data suggest that the fat-derived mesenchymal cells responded to soluble mediators from the disk. Future studies on intervertebral disk reconstruction could be based on our findings with fat-derived multipotential cells from the inguinal region of the rabbit that were co-cultured with disk tissue and may prove useful in tissue engineering strategies.

Médecine régénératrice du disque intervertébral

“…Plusieurs études précliniques ont permis de démontrer la capacité de cellules « régénératrices » (cellules du NP, cellules adultes, CSM non différenciées) à synthétiser une MEC cartilagineuse en l'absence de matériau [13,27,28]. Parfois imparfaites, notamment par l'absence d'uniformité entre les matériels et les méthodes, ces études précliniques ont malgré tout permis d'envisager la mise en oeuvre d'études cliniques.…”

Alginate cell encapsulation: new advances in reproduction and cartilage regenerative medicine

“…ADSCs regenerative therapy has also been implied for the treatment of IVD diseases (O'Halloran and Pandit 2007;Gaetani et al 2008;Helder et al 2007;Li et al 2005;Richardson et al 2006): in fact, chondrocytes could be retrieved, expanded in a 3D in vitro system consisting of alginate capsules, and finally re-implanted through a mini-invasive technique. Moreover it is now possible to regenerate or replace damaged IVDs through the implants of biodegradable scaffolds seeded with autologous ADSCs previously induced to differentiate to the chondrogenic phenotype (Helder et al 2007).…”

“…Moreover, in rabbits, the adipose tissue-derived cells co-cultured with Nucleus Polposus (NP) cells showed a significant increase in the expression of type II collagen and aggrecan genes, suggesting that ADSCs might respond to mediators contained in the IVD (Li et al 2005). The barium alginate encapsulation technique applied by Gaetani et al (2008) showed that ADSCs improve the quality of in vitro reconstructed NP tissue for the regeneration of the human degenerated IVD.…”

Demineralized bone matrix gelatin as scaffold for osteochondral tissue engineering

“…After the tissue was homogenized with a rotor-stator homogenizer, total RNA was extracted and purified using a RNeasy kit (Qiagen, Chatsworth, CA); purified RNA was subjected to real-time PCR (RT-PCR) analysis according to our published protocol [22]. The sequence of primers were as follows: for type I collagen (D49399): 5 0 -CGGTGGTTACGACTTTGGTT-3 0 , 3 0 -TCAGAGTGG-CATCGACTTCA-5 0 ; for type II collagen gene (D83228): 5 0 -AACA-CTGCCAACGTCCAGAT-3 0 , 3 0 -CTGCAGCACGGTATAGGTGA-5 0 ; for aggrecan gene (L38480): 5 0 -GAGGTCGTGGTGAAAGGTGT-3 0 , 3 0 -GTGTGGATGGGGTACCTGAC-5 0 .…”