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Three-dimensional lithographically defined organotypic tissue arrays for quantitative analysis of morphogenesis and neoplastic progression - PubMed

Three-dimensional lithographically defined organotypic tissue arrays for quantitative analysis of morphogenesis and neoplastic progression

Celeste M Nelson et al. Nat Protoc. 2008.

Abstract

Here, we describe a simple micromolding method to construct three-dimensional arrays of organotypic epithelial tissue structures that approximate in vivo histology. An elastomeric stamp containing an array of posts of defined geometry and spacing is used to mold microscale cavities into the surface of type I collagen gels. Epithelial cells are seeded into the cavities and covered with a second layer of collagen. The cells reorganize into hollow tissues corresponding to the geometry of the cavities. Patterned tissue arrays can be produced in 3-4 h and will undergo morphogenesis over the following 1-3 d. The protocol can easily be adapted to study a variety of tissues and aspects of normal and neoplastic development.

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Figures

Figure 1
Figure 1

Schematic of method to pattern multicellular tissues in micromolded gels.

Figure 2
Figure 2

Images of different stages of the patterning process. (a) PDMS stamp (inset: vertical section through one post); (b) molded collagen gel; (c) molded gel during addition of cells (note that cells are both in the wells and on top of the gel); (d) molded gel after washing away excess cells; (e) tubules; (f) branched tissues 24 h after addition of EGF to the sample; (g) one branched tissue stained for nuclei with Hoechst 33258; (h) frequency map depicting quantification of 50 branched tissues. Scale bars, 200 µm (a–f) and 50 µm (g,h).

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