Chitooligosaccharide (CHOS), a depolymerized chitosan, can be prepared via physical, chemical, and enzymatic hydrolysis, or a combination of these techniques. The superior properties of CHOS have attracted attention as alternative additives or bioactive compounds for various food and biomedical applications. To increase the bioactivities of a CHOS, its derivatives have been prepared via different methods and were characterized using various analytical methods including FTIR and NMR spectroscopy. CHOS derivatives such as carboxylated CHOS, quaternized CHOS, and others showed their potential as potent anti-inflammatory, anti-obesity, neuroprotective, and anti-cancer agents, which could further be used for human health benefits. Moreover, enhanced antibacterial and antioxidant bioactivities, especially for a CHOS-polyphenol conjugate, could play a profound role in shelf-life extension and the safety assurance of perishable foods via the inhibition of spoilage microorganisms and pathogens and lipid oxidation. Also, the effectiveness of CHOS derivatives for shelf-life extension can be augmented when used in combination with other preservative technologies. Therefore, this review provides an overview of the production of a CHOS and its derivatives, as well as their potential applications in food as either additives or nutraceuticals. Furthermore, it revisits recent advancements in translational research and in vivo studies on CHOS and its derivatives in the medical-related field.