Chondrogenesis in a hyaluronic acid scaffold: comparison between chondrocytes and MSC from bone marrow and adipose tissue

Abstract

Treatment of focal lesions of the articular cartilage of the knee using chondrocytes in a hyaluronic acid (HA) scaffold is already being investigated in clinical trials. An alternative may be to use mesenchymal stem cells (MSC). We have compared articular chondrocytes with MSC from human bone marrow (BM) and adipose tissue (AT), all cultured in HA scaffolds, for their ability to express genes and synthesize proteins associated with chondrogenesis. The cells were expanded in monolayer cultures. After seeding into the scaffold, the chondrocytes were maintained in medium, while the two MSC populations were given a chondrogenic differentiation medium. Chondrogenesis was assessed by real-time RT-PCR for chondrocyte-associated genes, by immunohistochemistry and by ELISA for collagens in the supernatant. Redifferentiation of the dedifferentiated chondrocytes in the HA scaffold was shown by a modest increase in type II collagen mRNA (COL2A1) and reduction in COL1A1. BM-MSC expressed 600-fold higher levels of COL2A1 than chondrocytes after 3 weeks in the scaffold. The levels of aggrecan (AGC1) and COL1A1 were similar for chondrocyte and BM-MSC scaffold cultures, while COL10A1 was higher in the BM-MSC. AT-MSC expressed levels of COL2A1 and COL1A1 similar to chondrocytes, but less AGC1 and COL10A1. Surprisingly, little collagen II protein was observed in the scaffold. Instead, collagen II was found in the culture medium. Chondrogenesis in HA scaffolds was more efficient using BM-MSC than AT-MSC or chondrocytes. Some of the secreted collagen II escaped entrapment in the extracellular space and was detected in the culture medium.

Epigenetic and in vivo comparison of diverse MSC sources reveals an endochondral signature for human hematopoietic niche formation

“…24 Adipose tissue was obtained by liposuction from healthy female donors aged 18 to 39 years as described. 17,25 UC-derived MSCs and skinderived MSC were isolated as previously described. 18 Briefly, biopsy pieces were allowed to adhere to culture dishes (Corning) before cell culture medium was carefully added.…”

microRNA-140 TargetsRALAand Regulates Chondrogenic Differentiation of Human Mesenchymal Stem Cells by Translational Enhancement ofSOX9andACAN

“…The Regional Committee for Medical Research Ethics, Southern Norway, approved the study, and all donors gave written consent. BM-MSCs and ACs were isolated and cultured as described previously [16,17] and in Supplementary Material.…”

“…cDNA synthesis and RT-qPCR was performed following protocols from the manufacturer using the High Capacity cDNA Reverse Transcripton Kit (Applied Biosystems) [17]. Subsequently, RT-qPCR was performed using the Taqman Gene or MicroRNA Expression assays and Taqman Universal PCR master mix with the 7300 Real-Time RT Global miRNA profiling BM-MSCs were differentiated in alginate in chondrogenic medium as described in Supplementary Material.…”

Bone and Cartilage from Stem Cells: Growth Optimalization and Stabilization of Cell Phenotypes

“…Integrins are extracellular receptors conveying mechano-stimulation to the interior of the cell (Liu, Calderwood et al 2000;Weyts, Li et al 2002;Kapur, Baylink et al 2003;Gordeladze, Reseland et al 2009), and the use of RDG alginate scaffolds makes the addition of growth factors less critical for chondrocyte phenotype acquisition (as shown by transcriptome analyses). Hyalouronan (HYAFF-11 ® ) scaffolds have been shown to produce useful cells for osteochondral tissue replacement, provided that MSCs were applied instead of ASCs in the presence of TGF 1 (Loken, Jakobsen et al 2008;Jakobsen, Shahdadfar et al 2010). Finally, it should be mentioned that scaffold material (e.g.…”