Chitooligosaccharides, also known as chitosan oligomers or chitooligomers, are made up of chitosan with a degree of polymerization (DP) that is less than 20 and an average molecular weight (MW) that is lower than 3.9 kDa. COS can be produced through enzymatic conversions using chitinases, physical and chemical applications, or a combination of these strategies. COS is of significant interest for pharmacological and medical applications due to its increased water solubility and non-toxicity, with a wide range of bioactivities, including antibacterial, anti-inflammatory, anti-obesity, neuroprotective, anticancer, and antioxidant effects. This review aims to outline the recent advances and potential applications of COS in various diseases and conditions based on the available literature, mainly from preclinical research. The prospects of further in vivo studies and translational research on COS in the medical field are highlighted.