Fabrication and Characterization of Immature Porcine Cartilage-Derived Cell Biomembranes

(1) Background: Knee cartilage injury is at the top of the rising concerns among bone and joint disorder patients. Autologous chondrocyte implantation (ACI) is widely used to approach knee cartilage deterioration. Integrating autologous chondrocytes and periosteal patches aids in forming new cartilage-like tissue at the lesion area. This study uses a novel cell source from one-day-old porcine cartilage to fabricate a biomembrane as a substitute for periosteal membranes in cell implantation techniques for treating knee cartilage injuries. (2) Methods: Cells isolated from one-day-old porcine cartilage tissue were identified and assessed for their proliferation capability, differentiation ability, and membrane formation potential. The protein component of the biomembrane was also defined by proteomics. The cartilage repair ability was also confirmed using an in vitro transplantation model. (3) Results: Negative results for porcine infectious diseases are pivotal in selecting suitable piglets to provide cartilage tissue. The cells successfully obtained from one-day-old porcine cartilage exhibited stem-cell-like characteristics (CD34-, CD45-, CD90+, CD105+), including a high proliferation to 20 passages (doubling time: 1–2 days) and a capacity to differentiate into various cell types (osteogenesis, adipogenesis, and chondrogenesis). The stem cells were successfully applied in the fabrication of the biomembranes. The protein components of the biomembrane included an extracellular matrix and growth factors. The in vitro transplantation model showed that the biomembrane induced the repair ability of cartilage defects. (4) Conclusions: This study is the first to successfully harvest stem cells from one-day-old porcine cartilage for biomembrane fabrication for a knee cartilage injury therapeutic application.