Bardsley, K and Yang, Y and El Haj, AJ (2017) Fluorescent Labeling of Collagen Production by Cells for Noninvasive Imaging of Extracellular Matrix Deposition. Tissue Eng Part C Methods, 23 (4). 228 - 236. ISSN 1937-3392

[img] Text
K Bardsley - Fluorescent labelling of collagen production by cells for non-invasive....pdf - Accepted Version
Restricted to Repository staff only until 24 February 2018.

Download (152kB)
[img] Text
K Bardsley - Figure Legends.pdf - Accepted Version
Restricted to Repository staff only until 24 February 2018.

Download (159kB)
[img] Slideshow
K Bardsley - Figures.pptx - Accepted Version
Restricted to Repository staff only until 24 February 2018.

Download (6MB)

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.

Item Type: Article
Additional Information: Final publication is available from Mary Ann Liebert, Inc., publishers http://dx.doi.org/10.1089/ten.TEC.2017.0008
Subjects: Q Science > Q Science (General)
R Medicine > R Medicine (General)
?? collagen ??
?? extracellular matrix ??
?? fluorescence ??
?? noninvasive imaging ??
Divisions: Faculty of Medicine and Health Sciences > Institute for Science and Technology in Medicine
Related URLs:
Depositing User: Symplectic
Date Deposited: 22 May 2017 10:41
Last Modified: 22 May 2017 10:41
URI: http://eprints.keele.ac.uk/id/eprint/3486

Actions (login required)

View Item View Item