Cell Viability Assays for 3D Cellular Constructs

In vitro models fall short of replicating the complex in vivo processes including cell growth and differentiation. For many years, molecular biology research and drug development have relied on the use of cells grown within tissue culture dishes. These traditional in vitro two-dimensional (2D) cultures fail to recapitulate the 3D microenvironment of in vivo tissues. Due to inadequate surface topography, surface stiffness, cell-to-cell, and cell-to-ECM matrices, 2D cell culture systems are incapable of mimicking cell physiology seen in living healthy tissues. These factors can also place selective pressure on cells that substantially alter their molecular and phenotypic properties. With these disadvantages in mind, new and adaptive cell culture systems are necessary to recapitulate the cellular microenvironment in a more accurate manner for drug development, toxicity studies, drug delivery, and much more. Newly developed biofabrication technologies capable of creating 3D tissue constructs can open new opportunities for cell growth and developmental modeling. These constructs show great promise in representing an environment that allows cells to interact with other cells and their microenvironment in a much more physiologically accurate manner. When transitioning from 2D to 3D systems, there is the need to translate common cell viability analysis techniques from that of 2D cell culture to these 3D tissue constructs. Cell viability assays are critical in evaluating the health of cells in response to drug treatment or other stimuli to better understand how these factors effect the tissue constructs. As 3D cellular systems become the new standard in biomedical engineering, this chapter provides different assays used to assess cell viability qualitatively and quantitatively in 3D environments.