In vitro culture of primary or established cell lines is one of the leading techniques in many
areas of basic biological research. The use of pure or highly enriched cultures of specific cell types obtained
from different tissues and genetics backgrounds has greatly contributed to our current understanding
of normal and pathological cellular processes. Cells in culture are easily propagated generating an
almost endless source of material for experimentation. Besides, they can be manipulated to achieve gene
silencing, gene overexpression and genome editing turning possible the dissection of specific gene functions
and signaling pathways. However, monolayer and suspension cultures of cells do not reproduce the
cell type diversity, cell-cell contacts, cell-matrix interactions and differentiation pathways typical of the
three-dimensional environment of tissues and organs from where they were originated. Therefore, different
experimental animal models have been developed and applied to address these and other complex
issues in vivo. However, these systems are costly and time consuming. Most importantly the use of animals
in scientific research poses moral and ethical concerns facing a steadily increasing opposition from
different sectors of the society. Therefore, there is an urgent need for the development of alternative in
vitro experimental models that accurately reproduce the events observed in vivo to reduce the use of animals.
Organotypic cultures combine the flexibility of traditional culture systems with the possibility of
culturing different cell types in a 3D environment that reproduces both the structure and the physiology
of the parental organ. Here we present a summarized description of the use of epithelial organotypic for
the study of skin physiology, human papillomavirus biology and associated tumorigenesis.
In vivo three-dimensional velocity vector imaging and volumetric flow rate measurements
