Thanks to their high porosity and surface area, mesoporous bioactive glasses (MBGs) have gained significant interest in the field of medical applications, in particular, with regards to enhanced bioactive properties which facilitate bone regeneration. The aim of this article is to review the state of the art regarding the biocompatibility evaluation of MBGs and provide a discussion of the various approaches taken. The research was performed using PubMed database and covered articles published in the last five years. From a total of 91 articles, 63 were selected after analyzing them according to our inclusion and exclusion criteria. In vitro methodologies and techniques used for biocompatibility assessment were investigated. Among the biocompatibility assessment techniques, scanning electron microscopy (SEM) has been widely used to study cell morphology and adhesion. Viability and proliferation were assessed using different assays including cell counting and/or cell metabolic activity measurement. Finally, cell differentiation tests relied on the alkaline phosphatase assay; however, these were often complemented by specific bimolecular tests according to the exact application of the mesoporous bioactive glass. The standardization and validation of all tests performed for MBG cytocompatibility is a key aspect and crucial point and should be considered in order to avoid inconsistencies, bias between studies, and unnecessary consumption of time. Therefore, introducing standard tests would serve an important role in the future assessment and development of MBG materials.