Department of Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo.
Division of Tissue Engineering, The University of Tokyo Hospital.
Biomed Res. 2020;41(2):67-80. doi: 10.2220/biomedres.41.67.
Chondrocytes are utilized to cartilage regeneration by being harvested through enzymatic digestion and expanded by monolayer culture. However, these procedures will cause deterioration and dedifferentiation of the chondrocytes. In addition, scaffolds are often needed to provide the cartilage with mechanical strength and three-dimensional structures. We tried to use diced cartilage prepared using a micro-slicer without digestion, monolayer culture or scaffolds. In this study, an appropriate culture condition to induce the fusion of diced cartilage in vitro and cartilage regeneration in vitro and in vivo was determined to realize a scaffold-free cartilage regeneration. As a result, diced cartilages aggregated when they were cultured more than 5 weeks in the media containing 10% fetal bovine serum (FBS). Diced cartilage cultured for 7 weeks with the media containing 10%, followed by the culture with the media containing insulin-like growth factor-1 for 5 weeks in the ultralow attachment plate showed most prominent cartilage formation both in vitro and in vivo. The volume of regenerated cartilage was 2.14 times larger than that of the original cartilage. These results indicated that large regenerative cartilage from a small amount of cartilage was achieved without deterioration or dedifferentiation.
软骨细胞通过酶消化收获并通过单层培养进行扩增，用于软骨再生。然而，这些程序会导致软骨细胞的恶化和去分化。此外，通常需要支架为软骨提供机械强度和三维结构。我们尝试使用微切片器制备的未经消化、未经单层培养或无支架的软骨块。在这项研究中，确定了一种合适的培养条件，以诱导体外软骨块融合和体外及体内软骨再生，从而实现无支架的软骨再生。结果表明，当在含有 10%胎牛血清（FBS）的培养基中培养超过 5 周时，软骨块会聚集。在超低附着平板上用含有 10%的培养基培养 7 周，然后用含有胰岛素样生长因子-1 的培养基培养 5 周，在体外和体内都显示出最显著的软骨形成。再生软骨的体积比原始软骨大 2.14 倍。这些结果表明，在不恶化或去分化的情况下，从少量软骨中获得了大量的再生软骨。
