Abstract
Collagen- and fibrin-based gels are extensively used to study cell behaviour. However, 2D–3D and collagen-fibrin comparisons of gene expression, cell shape and mechanotransduction, with an in vivo reference, have not been reported. Here we compared chick tendon fibroblasts (CTFs) at three stages of embryonic development with CTFs cultured in collagen- or fibrin-based tissue engineered constructs (TECs). CTFs synthesised their own collagen matrix in fibrin-based TECs and better recapitulated the gene expression, collagen fibril alignment and cell shape seen in vivo. In contrast, cells in 3D collagen gels exhibited a 2D-like morphology and expressed fewer of the genes expressed in vivo. Analysis of YAP/TAZ target genes showed that collagen gels desensitise mechanotransduction pathways. In conclusion, gene expression and cell shape are similar on plastic and 3D collagen whereas cells in 3D fibrin have a shape and transcriptome better resembling the in vivo situation. Implications for wound healing are discussed.
Highlights
In this study we used embryonic chick tendon fibroblasts (CTFs) because they are highly motile in 2D culture, proliferate well, and can be isolated by clean dissection from chick embryos without the need for extensive handling procedures
Our first experiment was to identify genes expressed during tendon development in vivo and determine which of these genes are expressed by CTFs cultured in collagen-based and fibrin-based tissue engineered constructs (TECs)
RNA was isolated from CTFs cultured on 2D tissue culture plastic (2D), in restrained, attached fibrin gels, in taut tendon-like TECs (3D fibrin TEC), and in these fibrin-based TECs matured for 10 days in culture under tension[15]
Summary
In this study we used embryonic chick tendon fibroblasts (CTFs) because they are highly motile in 2D culture, proliferate well, and can be isolated by clean dissection from chick embryos without the need for extensive handling procedures. Three of the eight clusters contained genes that exhibited similar expression trends during the formation of both fibrin- and collagen-based TECs and in vivo tendon development (Supplementary Fig. 2b).
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