Cartilage has limited self-regenerative capacity. Tissue engineering can offer promising solutions for reconstruction of missing or damaged cartilage. A major challenge herein is to define an appropriate cell source that is capable of generating a stable and functional matrix. This study evaluated the performance of culture-expanded human chondrocytes from ear (EC), nose (NC) and articular joint (AC), as well as bone-marrow-derived and adipose-tissue-derived mesenchymal stem cells bothin vitroandin vivo. All cells (≥3 donors per source) were culture-expanded, encapsulated in alginate and cultured for 5 weeks. Subsequently, constructs were implanted subcutaneously for 8 additional weeks. Before and after implantation, glycosaminoglycan (GAG) and collagen content were measured using biochemical assays. Mechanical properties were determined using stress-strain-indentation tests. Hypertrophic differentiation was evaluated with qRT-PCR and subsequent endochondral ossification with histology. ACs had higher chondrogenic potentialin vitrothan the other cell sources, as assessed by gene expression and GAG content (p< 0.001). However, after implantation, ACs did not further increase their matrix. In contrast, ECs and NCs continued producing matrixin vivoleading to higher GAG content (p< 0.001) and elastic modulus. For NC-constructs, matrix-deposition was associated with the elastic modulus (R2= 0.477,p= 0.039). Although all cells – except ACs – expressed markers for hypertrophic differentiationin vitro, there was no bone formedin vivo. Our work shows that cartilage formation and functionality depends on the cell source used. ACs possess the highest chondrogenic capacityin vitro, while ECs and NCs are most potentin vivo, making them attractive cell sources for cartilage repair.