This research work explores the physicochemical characteristics and biological functions of polysaccharides extracted from four selected green seaweed species: Ulva lactuca (UL), Halimeda opuntia (HO), Caulerpa racemosa (CR), and Chaetomorpha antennina (CA). The extracts were investigated for cell-based bio-activities (i.e., cytotoxicity, cell proliferation and migration) using three cell lines (HDF, HaCaT, RAW264.7) reflecting cell models involved in wound healing, as well as anticoagulant activity. Ulvans from UL significantly increased HaCaT (at 0.06 μg/μL) and HDF (at 0.5 μg/μL) cell proliferation. In addition, extracts from CA showed the highest cell migration ability using HDF and HaCaT cells. UL (all fractions), HO-2A, CR-1B, CA-1A and CA-2B fractions improved phagocytosis. Furthermore, RAW264.7 cells treated with fraction CA-1A produced significantly more intracellular NO (pro-inflammatory) within 24 h compared to control (LPs). Green seaweed extracts CA-2A and UL-1A resulted in lower expression of the pro-inflammatory cytokine TNF-α in skin cells (HDF, HaCaT). Caulerpa cold-extracted polysaccharides possessed higher anticoagulant properties. The Ulva (1,4-linked α-L-rhamnose, β-D-glucuronic acid, L-iduronic acid) and CA extracts are promising sources of bioactive therapeutic agents. Our data provide useful insights into the possible biomedical benefits of selected polysaccharides mixtures (i.e., ulvan, sulfated or/and pyruvylated β-D-galactans, sulfated xyloarabinogalactan) for food, pharmaceutical, and biotechnological applications.