The layer-by-layer technique (LbL) using polysaccharides is receiving increasing interest as the preparation of nano- and micro- multilayer objects composed by natural materials is a promising alternative for actual systems. The use of bacterial polysaccharides and more particularly, dextrans and dextran derivatives, in LbL assemblies allows the simple formation of biodegradable and biocompatible multilayers with engineered functionalities. The choice of dextrans and derivatives along with the assembly conditions can indeed control internal structure and physical, chemical and biological properties. In this review, we describe the use of dextrans and dextran de- rivatives into multilayers presented in literature, from the method and principles to the key parameters that need to be considered in their formations. We present their valorisation into nanoobjects with specific and stimuli- responsive properties that are mainly exploited for biomedical applications as drug delivery devices.