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3D Organotypic Culture Model to Study Components of ERK Signaling.

Methods in molecular biology (Clifton, N.J.) (2016-12-08)
Athina-Myrto Chioni, Rabia Tayba Bajwa, Richard Grose
ABSTRACT

Organotypic models are 3D in vitro representations of an in vivo environment. Their complexity can range from an epidermal replica to the establishment of a cancer microenvironment. These models have been used for many years, in an attempt to mimic the structure and function of cells and tissues found inside the body. Methods for developing 3D organotypic models differ according to the tissue of interest and the experimental design. For example, cultures may be grown submerged in culture medium and or at an air-liquid interface. Our group is focusing on an air-liquid interface 3D organotypic model. These cultures are grown on a nylon membrane-covered metal grid with the cells embedded in a Collagen-Matrigel gel. This allows cells to grow in an air-liquid interface to enable diffusion and nourishment from the medium below. Subsequently, the organotypic cultures can be used for immunohistochemical staining of various components of ERK signaling, which is a key player in mediating communication between cells and their microenvironment.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Collagen Type I, rat tail
Sigma-Aldrich
Hanks′ Balanced Salt solution, 10 ×, Modified, without calcium, magnesium or sodium bicarbonate