Multi-potent stem cells derived from human adipose tissues, is a potential player in regenerative medicine; while very little is known about its interaction with cancer cells such as bone; raising the safety concern in their clinical utilization. In this context, the present study co-cultures human adipose derived stem cell (hASCs) with human osteosarcoma cells (SaOS 2) in a 3D micro-environment of cell-adhesive gellan gum- silk spongy-like hydrophilic networks. The hydrophilic cross-linked network of gellan gum is known to be supportive to maintain the phenotypes of osteosarcoma and hASCs cells. The incorporation of silk protein within this network affected the physical and mechanical properties of hydrogel; imparting the tuning ability to cell extracellular microenvironment (ECM). This investigation is focused on the delineating effect of hASCs on SaOS 2 cells within the tunable hydrogels. Cells exhibit aggregated to elongated phenotypes and stimulated the invasiveness through the production of Type I collagen based on cultured ECM. The migratory behavior of cells also detectably responds to the matrix composition; results in different cytoskeleton arrangements. Therefore, targeting ECM for cancer regulation and therapeutics development is envisioned from the study; however, further detail investigation is needed.