Recent advancements in research methodology signal a significant shift away from traditional animal models toward alternative approaches. While monolayer cell cultures have been standard, their limitations in replicating in vivo complexity have led to the rise of three-dimensional (3D) models. These innovative systems, including organoids, bioreactors, organ-on-a-chip, and bioprinting, overcome the drawbacks of traditional two-dimensional cultures, offering researchers the ability to recreate tissue heterogeneity, structure, and functions within a single dish.

Applications span regenerative medicine, drug discovery, precision medicine, cancer research, and gene expression studies. This shift not only revolutionizes cell culture technology but also provides ethical alternatives to animal testing. The comprehensive review presented here offers a comparative analysis between 2D and 3D cell cultures, delving into various 3D culture techniques. The discussion includes an exploration of their strengths and potential applications, highlighting the transformative impact these methodologies can have on both basic and specialized research. All these advancements align with the principles of the 3R approach (Replacement, Reduction, Refinement), contributing to more ethical and effective research practices.