Development of a Rotational Culture Method for Reconstruction of Mesodermal Tissue: A Preliminary Study

Cartilage regenerative medicine presents promising therapeutic options; however, mature cartilage tissue reconstruction remains challenging. Although techniques for expanding chondrocytes are clinically available, the production of mature articular and auricular cartilage tissues remains to be achieved. Similarly, although epidermal sheets from cell cultures have been clinically used for skin regeneration, dermal tissue reconstruction remains elusive. This study aimed to develop a rotational culture method to produce cartilage and dermal tissue without scaffold materials.

The articular and auricular cartilage and dermis were isolated from patients with polydactyly or accessory auricles. The chondrocytes and fibroblasts were transferred to culture vessels and subjected to different rotational culture conditions: lateral rotational to rapidly form sheets for articular cartilage, box rotational to thicken tissue for auricular cartilage, and bottom rotational to form large tissue sheets for dermal fibroblast and articular cartilage. Sections and cultured cells were subjected to hematoxylin-eosin, Alcian blue, and type I collagen staining.

The lateral rotational method produced articular cartilage-like sheets, as confirmed by Alcian blue staining, which identifies mucopolysaccharides that make up the cartilage matrix. The box rotational method produced auricular-like cartilage tissue from auricular chondrocytes, whereas the bottom rotational method produced dermal-like sheets expressing type I collagen from human skin fibroblasts. All tissues were produced without using scaffold materials through repeated cell seeding and culture rotation.

The tissues produced without scaffolds exhibited properties suitable for clinical applications, including elasticity, stretchability, and suturing capability, demonstrating a potentially promising breakthrough in cell manipulation technology for cartilage and dermis.