Annulus Fibrosus (AF) tissue integrity maintains intervertebral disc (IVD) structure, essential to spine mobility and shock absorption. However, this tissue, which confines nucleus pulposus (NP), has been poorly investigated, partially due to the lack of appropriate study models. This review provides a comprehensive analysis of AF in vitro models. By critically assessing the current AF in vitro models, this works thoroughly identifies key gaps in replicating the tissue's complex microenvironment. Finally, we outline the essential criteria for developing more accurate and reliable AF models, emphasizing the importance of biomaterial composition, architecture, and microenvironmental cues. By advancing in vitro models, we aim to deepen the understanding of AF failure mechanisms and support the development of novel therapeutic strategies for IVD herniation. Insights gained from this review may also have broader applications in regenerative medicine, particularly in the study and treatment of other connective tissue disorders. STATEMENT OF SIGNIFICANCE: This review evaluates the current in vitro models of the annulus fibrosus (AF), a key component of the intervertebral disc (IVD). By identifying gaps in these models, particularly in replicating tissue's complex microenvironment, we propose essential criteria for the development of more accurate AF models, to better understand the pathomechanisms and potentially aid the development of therapeutic approaches for spinal disorders. The findings also extend to broader studies of musculoskeletal tissue disorders in the context of regenerative medicine, appealing to a diverse biomedical research readership.