Tissue Engineering and Cell-Populated Collagen Matrices - Methods in Molecular Biology: Extracellular Matrix Protocols - page 274

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Tissue Engineering
and Cell-Populated Collagen Matrices

Paul D. Kemp

1. Introduction
Tissue engineering seeks to produce living, three-dimensional cellular
constructs that can be used as clinical replacements of damaged tissues and
organs as well as research tools to study cell and matrix interactions that
occur in higher-order systems. To organize the cells into a three-dimensional
structure in vitro, a provisional extracellular matrix support is required. The
two main methods to achieve this are a) to culture the stromal cells on a
three-dimensional synthetic meshwork, or else b) embed the cells within a
three-dimensional type I collagen lattice. The contracted collagen lattice can
be used for a variety of practical applications including the support of epithe-
lial growth and differentiation in order to produce a skin replacement (1-5) .
However, this model system can also be exploited for experiments to
study cell-matrix interactions such as the influence of tension on cell
phenotype (6) .

2. Materials
2.1. Production of the Collagen Solution
1. PBS:Water: Mix one part phosphate-buffered saline with 2 parts of purified

water. Filter sterilize the solution through an appropriate 0.22- µ m filter into a

sterile, screwcap storage vessel(s). Store the solution at 2-8 ° C before use (stable

for 1 yr).

2. 0.5 M Acetic Acid: Carefully make up 286 mL of glacial acetic acid to 10 L with

purified water. Filter sterilize the solution through an appropriate 0.22 m filter
into a sterile, screwcap storage vessel(s). Store the solution at 2-8 ° C before use

(stable for 1 yr).