Chondrocyte-based strategies are widely used for cartilage repair and show promising results in the treatment of articular cartilage damage and osteoarthritis. In cartilage repair approaches that involve the implantation of autologous chondrocytes or tissue-engineered scaffolds, it is essential to obtain significant amounts of viable cells. Nasal chondrocytes serve as a clinically valuable source of cells for cartilage regeneration, and their isolation is a critical step in cartilage tissue engineering. This chapter outlines an optimized protocol for obtaining nasal chondrocytes from neonatal rats using a Leica VT1200 vibratome to obtain nasal cartilage fragments. Enzymatic isolation of nasal chondrocytes and their subsequent in vitro culture highlight the importance of the stem and progenitor cell niche in supporting the further culture and differentiation of these cells for effective cartilage regeneration.