Three‐dimensional organization of dermal fibroblasts by macromass culture

Abstract

The three-dimensional organization of cells by high-cell-seeding-density culture, termed 'macromass culture', is described. By macromass culture, dermal fibroblasts can be made to organize themselves into a unified three-dimensional form without the aid of a scaffold, and macroscopic constructs, named macromasses, can be made wholly from cells. The sole factor causing three-dimensional organization is culture of cells at high cell seeding density per unit area. No scaffold or extraneous matrix is used for the generation of macromasses; they are of completely cellular origin. No other agents or external influences such as tissue-inducing chemicals, tissue-inducing growth factors, substratum with special properties, rotational culture, centrifugation etc. are employed for macromass formation, and all seeded cells become part of the cohesive construct. These three-dimensional constructs have the potential for use as in vitro tissue analogues, and a possible application for in vitro cytotoxicity testing is demonstrated.

A novel dermal tissue construct: Development and in vitro characterization

“…A higher density of cells in a skin substitute is expected to result in more efficient tissue regeneration 16. In the creation of the dermal construct, this has been achieved by utilizing the multilayered tissue‐like sheet generated by macromass culture,9 in which cells are present at a high density. Since the macromass tissue‐like sheet by itself is contractile and to improve mechanical strength of the dermal construct, in this work, the macromass sheet has been mounted onto a chitosan sponge.…”

“…Previously, the formation of a three‐dimensional multilayered sheet of dermal fibroblasts using high density culture has been described 9. These sheets were contractile in nature and could reduce to a very small size, not useful for coverage of a wound area.…”