Abstract
Collagen is a widely investigated extracellular matrix material with extensive potentials in the field of tissue engineering. This protocol describes a method to prepare reconstituted collagen that can be ready-to-use, storable and suitable for further in vitro and in vivo investigations. Type I collagen was extracted from rat tail tendons and processed in acetic acid solution to obtain sterile soluble collagen. At first, crude collagen was dissolved in acetic acid, then frozen at −20 °C and lyophilized to obtain a sponge, which could be stored at −80 °C. Lyophilized collagen was then dispersed in acetic acid to obtain a sterile solution of collagen at targeted concentrations. The whole low-cost process from the extraction to the final sterile solution takes around 2–3 weeks. The collagen solution, once neutralized, has the potential to be used to produce gels or scaffolds, to deposit thin films on supports and to develop drug delivery systems.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Berglund, J.D., Mohseni, M.M., Nerem, R.M. & Sambanis, A. A biological hybrid model for collagen-based tissue engineered vascular constructs. Biomaterials 24, 1241–1254 (2003).
Purna, S.K. & Babu, M. Collagen based dressings––a review. Burns 26, 54–62 (2000).
Wallace, D.G. & Rosenblatt, J. Collagen gel systems for sustained delivery and tissue engineering. Adv. Drug Deliv. Rev. 55, 1631–1649 (2003).
Sionkowska, A. & Kaminska, A. Thermal helix-coil transition in UV irradiated collagen from rat tail tendon. Int. J. Biol. Macromol. 24, 337–340 (1999).
Steven, F.S. & Tristram, G.R. The presence of non-protein nitrogen in acetic acid-soluble calf-skin collagen. Biochem. J. 83, 240–244 (1962).
Silver, F.H. & Trelstad, R.L. Type I collagen in solution. Structure and properties of fibril fragments. J. Biol. Chem. 255, 9427–9433 (1980).
Wood, G.C. The heterogeneity of collagen solutions and its effect on fibril formation. Biochem. J. 84, 429–435 (1962).
Gross, J., Highberger, J.H. & Schmitt, F.O. Extraction of collagen from connective tissue by neutral salt solutions. Proc. Natl. Acad. Sci. USA 41, 1–7 (1955).
Habermehl, J. et al. Preparation of a ready-to-use, stockable and reconstituted collagen. Macromol. Biosci. 5, 821–828 (2005).
Boccafoschi, F., Habermehl, J., Vesentini, S. & Mantovani, D. Biological performances of collagen-based scaffolds for vascular tissue engineering. Biomaterials 26, 7410–7417 (2005).
Sionkowska, A. Flash photolysis and pulse radiolysis studies on collagen type I in acetic acid solution. J. Photochem. Photobiol. B 84, 38–45 (2006).
Elsdale, T. & Bard, J. Collagen substrata for studies on cell behavior. J. Cell. Biol. 54, 626–637 (1972).
Bell, E., Ivarsson, B. & Merrill, C. Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. Proc. Natl. Acad. Sci. USA 76, 1274–1278 (1979).
Weinberg, C.B. & Bell, E. A blood vessel model constructed from collagen and cultured vascular cells. Science 231, 397–400 (1986).
Deyl, Z., Miksik, I. & Eckhardt, A. Preparative procedures and purity assessment of collagen proteins. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 790, 245–275 (2003).
Lee, C.H., Singla, A. & Lee, Y. Biomedical applications of collagen. Int. J. Pharm. 221, 1–22 (2001).
Acknowledgements
This work was partially supported by the Natural Science and Engineering Research Council of Canada, the Canadian Institute for Health Research and the Quebec City University Hospital Research Center.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Rights and permissions
About this article
Cite this article
Rajan, N., Habermehl, J., Coté, MF. et al. Preparation of ready-to-use, storable and reconstituted type I collagen from rat tail tendon for tissue engineering applications. Nat Protoc 1, 2753–2758 (2006). https://doi.org/10.1038/nprot.2006.430
Published:
Issue Date:
DOI: https://doi.org/10.1038/nprot.2006.430
This article is cited by
-
Self-Assembling Peptide Nanofibers Anchored Parathyroid Hormone Derivative for Bone Tissue Engineering
Advanced Fiber Materials (2024)
-
Extraction and Characterization of Human Adipose Tissue-Derived Collagen: Toward Xeno-Free Tissue Engineering
Tissue Engineering and Regenerative Medicine (2024)
-
An Injectable In Situ Forming Collagen/Alginate/CaSO4 Composite Hydrogel for Tissue Engineering Applications: Optimization, Characterization and In Vitro Assessments
Arabian Journal for Science and Engineering (2024)
-
Keratose Hydrogel Drives Differentiation of Cardiac Vascular Smooth Muscle Progenitor Cells: Implications in Ischemic Treatment
Stem Cell Reviews and Reports (2023)
-
Methodological Insights on Morphometric Comparison of Collagen-Type-1 and Polystyrene Grown Malignant Glioma Cells Upon Chemical Stress Induction
Proceedings of the National Academy of Sciences, India Section B: Biological Sciences (2023)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.