For more than 70 years, acoustic waves have been used to screen, diagnose, and treat patients in hundreds of medical devices. The biocompatible nature of acoustic waves, their non-invasive and contactless operation, and their compatibility with wide visualization techniques are just a few of the many features that lead to the clinical success of sound-powered devices. The development of microelectromechanical systems and fabrication technologies in the past two decades reignited the spark of acoustics in the discovery of unique microscale bio applications. Acoustofluidics, the combination of acoustic waves and fluid mechanics in the nano and micro-realm, allowed researchers to access high-resolution and controllable manipulation and sensing tools for particle separation, isolation and enrichment, patterning of cells and bioparticles, fluid handling, and point of care biosensing strategies. This versatility and attractiveness of acoustofluidics have led to the rapid expansion of platforms and methods, making it also challenging for users to select the best acoustic technology. Depending on the setup, acoustic devices can offer a diverse level of biocompatibility, throughput, versatility, and sensitivity, where each of these considerations can become the design priority based on the application. In this paper, we aim to overview the recent advancements of acoustofluidics in the multifaceted fields of regenerative medicine, therapeutic development, and diagnosis and provide researchers with the necessary information needed to choose the best-suited acoustic technology for their application. Moreover, the effect of acoustofluidic systems on phenotypic behavior of living organisms are investigated. The review starts with a brief explanation of acoustofluidic principles, the different working mechanisms, and the advantages or challenges of commonly used platforms based on the state-of-the-art design features of acoustofluidic technologies. Finally, we present an outlook of potential trends, the areas to be explored, and the challenges that need to be overcome in developing acoustofluidic platforms that can echo the clinical success of conventional ultrasound-based devices.