Macromolecularly crowded in vitro microenvironments accelerate the production of extracellular matrix-rich supramolecular assemblies

• Published in: Scientific reports Vol. 5; no. 1; p. 8729
• Main Authors: Kumar, Pramod, Satyam, Abhigyan, Fan, Xingliang, Collin, Estelle, Rochev, Yury, Rodriguez, Brian J., Gorelov, Alexander, Dillon, Simon, Joshi, Lokesh, Raghunath, Michael, Pandit, Abhay, Zeugolis, Dimitrios I.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 04.03.2015; Nature Publishing Group
• Subjects: 101/58; 13/106; 13/107; 13/51; 14/19; 14/3; 14/35; 38; 38/88; 49; 49/79; 631/1647/2204; 631/61/2035; 631/80/83; 692/308/575; 82; 82/29; Animals; Animals, Newborn; Cattle; Cell culture; Cell Culture Techniques; Cell Physiological Phenomena; Cells, Cultured; Cellular Microenvironment - drug effects; Cellular Microenvironment - physiology; Collagen - metabolism; Copolymers; Cornea; Cornea - cytology; Culture Media - chemistry; Culture Media - pharmacology; Electrophoresis, Polyacrylamide Gel; Extracellular matrix; Extracellular Matrix - metabolism; Fibroblasts - cytology; Fibroblasts - drug effects; Fibroblasts - metabolism; Humanities and Social Sciences; Humans; Macromolecular Substances - metabolism; Macromolecules; Matrix Metalloproteinase 2 - metabolism; Microenvironments; Microscopy, Atomic Force; Microscopy, Fluorescence; Microscopy, Phase-Contrast; Models, Biological; multidisciplinary; Procollagen - metabolism; Science; Self-assembly; Serum - chemistry; Temperature effects; Time-Lapse Imaging; Tissue engineering; Tissue Engineering - methods
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep08729

Therapeutic strategies based on the principles of tissue engineering by self-assembly put forward the notion that functional regeneration can be achieved by utilising the inherent capacity of cells to create highly sophisticated supramolecular assemblies. However, in dilute ex vivo microenvironments, prolonged culture time is required to develop an extracellular matrix-rich implantable device. Herein, we assessed the influence of macromolecular crowding, a biophysical phenomenon that regulates intra- and extra-cellular activities in multicellular organisms, in human corneal fibroblast culture. In the presence of macromolecules, abundant extracellular matrix deposition was evidenced as fast as 48 h in culture, even at low serum concentration. Temperature responsive copolymers allowed the detachment of dense and cohesive supramolecularly assembled living substitutes within 6 days in culture. Morphological, histological, gene and protein analysis assays demonstrated maintenance of tissue-specific function. Macromolecular crowding opens new avenues for a more rational design in engineering of clinically relevant tissue modules in vitro .