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Tendons are able to undergo repeated cyclic loading in vivo without permanent deformation or mechanical failure. However, diseased, traumatised and decellularised tendons gradually lose the ability to resist load and fail because of creep deformation. The molecular basis of the mechanical properties of tendon and how cells establish and maintain these properties is poorly understood. New knowledge in this area is required to develop novel medical strategies to improve tendon repair and regeneration. Recent advances in tissue bioengineering have led to the formation of fibrin-based tendon-like tissue (‘tendon constructs’) that display the mechanical properties and ultrastructure of embryonic tendon. This thesis presents the characterisation of the tendon constructs derived from primary fibroblasts to understand the relationship between the cells and matrix during tissue development, and to establish the standard of in vitro engineered tendons. These findings facilitated protocol development to engineer human tendon-like tissue derived from stem cells. Novel findings of constructs formed from differentiated human pluripotent stem cells in feeder and feeder-free systems are presented. Fibrin gels were seeded with human dermal fibroblasts (HDF), chick tendon fibroblasts (CTF), MAN5 (Manchester, embryonic stem) cells, human embryonic stem cells (HuES7) and induced pluripotent stem cells (iPS). The gels were cultured until isometric tendon-like constructs were formed (T0) or continued for four or ten days post-formation. The mechanical properties, histology and gene expression of the constructs were analysed and compared between the constructs seeded with the aforementioned cell types. Varying the initial cell number (tested in CTF-seeded fibrin and collagen based constructs) significantly affected the final cell count and the mechanical properties of the constructs differentially at T0 and T10. A non-linear relationship exists between the initial and final cell number, and, between the initial cell number and mechanical properties. However, the results showed that cell number impacted cell-matrix stabilisation as strength per se was strongly dependent on initial cell number. Collagen-based constructs showed a significantly lower stiffness compared with fibrin-based constructs at T0 and T10. The stem cells and primary cells reproducibly underwent morphogenesis to form a 3D tissue similar to embryonic tendon in vivo expressing ECM markers such as collagens type I and III. The tissue also exhibited the ultrastructural characteristics and biomechanical profile of immature tendons. RNA seq and qPCR results demonstrated the upregulation of tendon-specific genes. Tendon-like tissue generated from human stem cells and HDFs in vitro has the potential to replace functional tissue lost through disease and to advance the understanding of the molecular basis of human tenogenesis.