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Enzymatically triggered peptide hydrogels for 3D cell encapsulation and culture

Szkolar, Laura; Guilbaud, Jean-Baptiste; Miller, Aline F; Gough, Julie E; Saiani, Alberto

Journal of Peptide Science. 2014;20(7):578-584.

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Abstract

We have investigated the possibility of using enzymatically triggered peptide hydrogels for the encapsulation and culture of cells. Based on recent work done on the enzymatically triggered gelation of FEFK (F, phenylalanine; E, glutamic acid; K, lysine) using thermolysin, a protease enzyme from Bacillus Thermoproteolyticus Rokko, we have investigated the possibility of using this gelation triggering mechanism to encapsulate cells within a 3D hydrogel matrix. First, the properties of enzymatically triggered hydrogels prepared in phosphate buffer solution were investigated and compared with the properties of hydrogels prepared in HPLC grade water from our previous work. We showed that the use of phosphate buffer solution allowed the production of hydrogels with very high shear moduli (>1MPa). The gelation kinetics was also investigated, and the mechanical properties of the system were shown to closely follow the synthesis of the octapeptide by the enzyme through reverse hydrolysis. In a second phase, we developed, on the basis of information acquired, a facile protocol for the encapsulation of cells and plating of the hydrogel. Human dermal fibroblasts were then used to exemplify the use of these materials. FEFEFKFK octapeptide hydrogels prepared under the same conditions and with the same mechanical properties were used as a control. We showed that no significant differences were observed between the two systems and that after a decrease in cell number on day 1, cells start to proliferate. After 5days of culture, the cells can be seen to start to adopt a stretched morphology typical of fibroblasts. The results clearly show that the protocol developed minimises the potential detrimental effect that thermolysin can have on the cells and that these enzymatically triggered hydrogels can be used for the 3D encapsulation and culture of cells. Copyright (c) 2014 European Peptide Society and John Wiley & Sons, Ltd.

Bibliographic metadata

Type of resource:
Content type:
Publication status:
Accepted
Publication type:
Publication form:
Published date:
ISSN:
Volume:
20
Issue:
7
Start page:
578
End page:
584
Total:
7
Pagination:
578-584
Digital Object Identifier:
10.1002/psc.2666
ISI Accession Number:
WOS:000337640600015
Related website(s):
  • Related website <Go to ISI>://WOS:000337640600015
General notes:
  • Times Cited: 1 SI
Attached files embargo period:
Immediate release
Attached files release date:
21st January, 2015
Access state:
Active

Institutional metadata

University researcher(s):

Record metadata

Manchester eScholar ID:
uk-ac-man-scw:248661
Created by:
Saiani, Alberto
Created:
21st January, 2015, 14:32:42
Last modified by:
Saiani, Alberto
Last modified:
21st January, 2015, 20:38:10

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