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A new procedure for rapid, high yield purification of Type I collagen for tissue engineering

Paper ID Volume ID Publish Year Pages File Format Full-Text
35088 45074 2009 13 PDF Available
Title
A new procedure for rapid, high yield purification of Type I collagen for tissue engineering
Abstract

Collagen, which is one of the most abundant proteins in mammalian proteomes, is of significant medical and also biotechnological importance. In tissue engineering, collagen is used as natural matrix to facilitate growth of mammalian cells in 3D-structures. Here, large amounts of highly purified Type I collagen have been obtained from rat tail tendon by a two-step purification, suitable for industrial up-scaling, involving extraction with 9 M urea followed by Superose 12 chromatography. Mass spectrometry identified only collagen Type I peptides indicating that the extracted collagen was homogeneous. Furthermore, a substantial amount of post-translational modifications were identified, including the presence of hydroxylated proline at the X-position in the G-X-Y repeats, which give new insight into collagen structure in vivo. The purified collagen was renatured quantitatively to form triple-helices by dialysis against water, as judged by UV-circular dichroism. This urea-extracted collagen (UC) was tested for its suitability as a matrix for growth of mammalian cells in tissue engineering. Cultures of a fibroblast cell line grown on urea-extracted collagen showed a higher motility and reduced stress levels than those grown on acetic acid-extracted collagen as determined by morphological studies and transcriptional analysis of selected marker genes using real time-PCR. These results indicate the suitability of the urea-extracted collagen obtained for biotechnological applications and tissue engineering.

Keywords
Type I collagen; Urea-extraction; Mass spectrometry; Self-assembly; Cell culture; Tissue engineering
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A new procedure for rapid, high yield purification of Type I collagen for tissue engineering
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 44, Issue 11, November 2009, Pages 1200–1212
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us