With the advancement of medical technology, the previous biomedical material platforms have been unable to meet the increasingly diverse application requirements, and the emergence of poly(2-oxazoline)s provides an opportunity to develop the next generation of biomedical materials. The highly tunable structure and function of poly(2-oxazoline)s, with excellent physical and biological properties, have shown great potential for application in the initial exploratory work. Currently, there are a lot of applied studies related to the water solubility, invisibility, and thermoresponsive of poly(2-oxazoline)s, mainly focused on drug delivery, protein modifcation, gene carriers, anti-fouling interface, cell sheet engineering, and hydrogel. This paper describes the preparation and physicochemical properties of poly(2-oxazoline)s and reviews the recent applications of poly(2-oxazoline)s in the biomedical feld.