Tissue regeneration using in-vitro scaffold becomes a vital mean to mimic the in-vivo counterpart due to the insufficiency of animal models to predict the applicability of drug and other physiological behavior. Three-dimensional (3D) bio-printing is an emerging technology to reproduce living tissue through controlled allocation of biomaterial and cell. Due to its bio-compatibility, natural hydrogels are commonly considered as the scaffold material in bio-printing process. However, repeatable scaffold structure with good printability and shape fidelity is a challenge with hydrogel material due to weak bonding in polymer chain. Additionally, there are intrinsic limitations for bio-printing of hydrogels due to limited cell proliferation and colonization while cells are immobilized within hydrogels and don’t spread, stretch and migrate to generate new tissue. The goal of this research is to develop a bio-ink suitable for extrusion-based bio-printing process to construct 3D scaffold. In this research, a novel hybrid hydrogel, is designed and systematic quantitative characterization are conducted to validate its printability, shape fidelity and cell viability. The outcomes are measured and quantified which demonstrate the favorable printability and shape fidelity of our proposed material. The research focuses on factors associated with pre-printing, printing and post-printing behavior of bio-ink and their biology. With the proposed hybrid hydrogel, 2 cm tall acellular 3D scaffold is fabricated with proper shape fidelity. Cell viability of the proposed material are tested with multiple cell lines i.e. BxPC3, prostate stem cancer cell, HEK 293, and Porc1 cell and about 90% viability after 15-day incubation have been achieved. The designed hybrid hydrogel demonstrate excellent behavior as bio-ink for bio-printing process which can reproduce scaffold with proper printability, shape fidelity and higher cell survivability. Additionally, the outlined characterization techniques proposed here open-up a novel avenue for quantifiable bio-ink assessment framework in lieu of their qualitative evaluation.