The central nervous system (CNS) is highly sensitive to inflammatory processes, injuries, and trauma. Given its limited regenerative capacity, the CNS has developed a unique response to injury, whose major part is formation of a glial scar. The glial scar, a heterogeneous structure, contains multiple cell types, with microglia and astrocytes being particularly important. With their specific characteristics, these cells maintain homeostasis in a healthy CNS, while in case of injury they promote its recovery, which is limited. Microglia, located within the parenchyma, represent the primary immune guardians of this system. They detect the smallest changes through numerous surface receptors, quickly initiating an inflammatory response. Their specific amoeboid shape allows them to perform phagocytosis, which is crucial for maintaining CNS balance. Astrocytes, as heterogeneous cells, perform a variety of functions, including ionic homeostasis and the blood-brain barrier (BBB) maintenance, neural networks build up, synapses shaping, and neurotransmitters regulation. In the case of CNS injury, microglia and astrocytes undergo a series of morphological and functional changes, resulting in the formation of a dense astrocytic network around the lesion, as well as enhanced removal of cellular waste by microglia. The dynamic communication between microglia and astrocytes is fundamental to create a stable and functional scar that limits the spread of inflammation and promotes the regenerative process. Although the glial scar is essential for protecting healthy tissue, certain components, such as chondroitin sulfate proteoglycans (CSPGs) and collagen I, inhibit axonal growth and obstruct CNS recovery. Limited regeneration contributes to the development of neurodegenerative diseases; therefore, understanding the process of glial scar formation, with a focus on microglia-astrocyte interaction, is crucial for developing new therapies.