In vitro synthesis of native, fibrous long spacing and segmental long spacing collagen

• Published in: Journal of visualized experiments no. 67; p. e4417
• Main Authors: Loo, Richard W, Goh, Jane Betty, Cheng, Calvin C H, Su, Ning, Goh, M Cynthia
• Format: Journal Article
• Language: English
• Published: United States MyJove Corporation 20.09.2012
• Subjects: Adenosine Triphosphate - chemistry; Bioengineering; Collagen Type I - chemistry; Fibrillar Collagens - chemical synthesis; Fibrillar Collagens - chemistry; Fibrillar Collagens - ultrastructure; Microscopy, Atomic Force - methods; Orosomucoid - chemistry
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/4417

Collagen fibrils are present in the extracellular matrix of animal tissue to provide structural scaffolding and mechanical strength. These native collagen fibrils have a characteristic banding periodicity of ~67 nm and are formed in vivo through the hierarchical assembly of Type I collagen monomers, which are 300 nm in length and 1.4 nm in diameter. In vitro, by varying the conditions to which the monomer building blocks are exposed, unique structures ranging in length scales up to 50 microns can be constructed, including not only native type fibrils, but also fibrous long spacing and segmental long spacing collagen. Herein, we present procedures for forming the three different collagen structures from a common commercially available collagen monomer. Using the protocols that we and others have published in the past to make these three types typically lead to mixtures of structures. In particular, unbanded fibrils were commonly found when making native collagen, and native fibrils were often present when making fibrous long spacing collagen. These new procedures have the advantage of producing the desired collagen fibril type almost exclusively. The formation of the desired structures is verified by imaging using an atomic force microscope.