Fluorescent Labeling of Collagen Production by Cells for Noninvasive Imaging of Extracellular Matrix Deposition.

Yang

doi:10.1089/ten.TEC.2017.0008

Fluorescent Labeling of Collagen Production by Cells for Noninvasive Imaging of Extracellular Matrix Deposition.

Yang

Authors

Ying Yang y.yang@keele.ac.uk

Abstract

Extracellular matrix (ECM) is an essential component of tissues and provides both integrity and biological cues for cells. Collagen is one of the major proteins found within the ECM and therefore is an essential component of all engineered tissues. Therefore, in this article, we present a method for the online real-time monitoring of collagen deposition in three-dimensional engineered constructs. This method revolves around modification of collagen through the addition of azide-L-proline to cell culture media. The incorporation of azide-L-proline into the neocollagen produced by cells can then be detected by reaction with 10?mM of a Click-IT Alexa Fluor 488 DIBO Alkyne. The reaction was shown as being specific to the collagen as little background staining was observed in cultures, which did not contain the modified proline, and the staining was also depleted after treatment with collagenase and colocalization of collagen type I staining by immunochemistry assay. Real-time online staining of collagen deposition was observed under different culture conditions without affecting proliferation. Collagen deposition was observed to be increased under mechanical stimulation; however, the localization varied across stimulation regimes. This is a new technique for real-time monitoring of cell-produced collagen and will be a valuable addition to the tissue engineering field.

Acceptance Date	Jan 6, 2017
Publication Date	Feb 24, 2017
Journal	Tissue Eng Part C Methods
Print ISSN	1937-3384
Publisher	Mary Ann Liebert
Pages	228 - 236
DOI	https://doi.org/10.1089/ten.TEC.2017.0008
Keywords	Fluorescence, Non-invasive Imaging, Collagen, Extracellular Matrix
Publisher URL	http://online.liebertpub.com/doi/abs/10.1089/ten.TEC.2017.0008#utm_source=JA&utm_medium=email&utm_campaign=tec