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Traditional cell culture techniques and protocols are performed on two-dimensional (2D) surfaces. While the 2D method is accessible and convenient, several well-documented artifacts can arise from studying cells cultured under 2D conditions. Advanced cell culture methods and protocols have been developed to culture cells in 3D environments in vitro. Cell culture scaffold, typically composed of biopolymers that are arranged to mimic the physiological extracellular matrix (ECM), is developed for complex tissue engineering projects, like developing artificial organs, establishing bioreactors to mass-produce cellular products, or developing lab-grown or cultured meat.