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Durable coating with extracellular matrix for cell culture through bio orthogonal functionalization of a extracellular matrix

Abstract

During a joint research project scientists at the University of Stuttgart and the University of Konstanz developed an ECM with functional groups (clickECM) which is covalently and thus firmly bonded to surfaces via bioorthogonal ligation reactions.

The fixed bonding obtained through the click reaction also ensures that the clickECM coating, e.g. when used for cell culture vessels, withstands extensive washing. The coating is highly stable and can be produced economically without any costly purification or isolation processes. The clickECM only differs from a natural ECM in terms of the introduced functional group.

Background

An extracellular matrix forms the natural environment of cells in biological tissue. It is involved in various cellular processes, for instance signal transmission, reproduction and adherence of the cells to surfaces. The composition of an ECM largely depends on the cell type it was secreted from. These tissue-specific matrices differ in the composition and proportional quantity of their biomolecules. Thus, depending on the origin of an ECM, it can e.g. either promote or maintain the differentiation of stem cells. This makes it an interesting potential candidate for use in special cell cultures.
The clickECM coating according to the invention allows for the simple production of a stable and application-oriented substrate, consisting of functionalized ECM, e.g. for cell culture vessels.

Problem

Conventional cell culture dishes are made of polystyrene and undergo plasma treatment to promote cell adhesion and growth. For primary cell growth, ECM proteins are normally physisorbed on the surface, supported by previous plasma technology, if required. Methods for equipping surfaces with proteins via covalent bonds are used in current state-of-the-art, but are not suitable for a coating with complex matrices. So far, coatings with complex ECM have only been performed via physisorption. Although the results obtained confirm the high biological activity of the ECM, the technology can only be used with certain materials. A particular disadvantage of such applied ECM coatings is their lack of stability, as the biomolecules only adhere to the surface via physisorption. These coatings can be washed off the substrate quickly and easily or might dissolve in other ways.

Solution

Until now it was very difficult to coat an artificial material with the components of the extracellular matrix. In the invention, a small, non-toxic reactive molecule is incorporated into the extracellular matrix. This enables the biological extracellular matrix to be bonded chemically to the surface of the cell culture dish. The coating process according to the invention not only promotes adhesion of the cells, but also strengthens cell growth. The clickECM only distinguishes itself from a natural ECM through its functional groups. With the help of the click-groups introduced, the clickECM can be stably and site-specifically immobilized on the surface of the likewise functionalized carrier. The fixed bonding obtained through the click reaction also ensures that the clickECM coating withstands extensive washing. A main advantage of clickECM is that it can be used to coat various new, innovative materials.

In-vitro production of the clickECM
In-vitro production of the clickECM (Photo: Fraunhofer IGB, Stuttgart)

Advantages

  • Coating with complex ECM possible
  • High stability of coating
  • Strong adhesion to various materials
  • Increased cell growth
  • Cost-effective production, as there is no elaborate purification, isolation or sterilization required

Application

Coating for cell culture vessels

Exposé
Contact
Dr. Frank Schlotter
TLB GmbH
Ettlinger Straße 25
76137 Karlsruhe | Germany
Phone +49 721-79004-0
schlotter(at)tlb.de | www.tlb.de
Development Status
Late basic research / TRL2
Patent Situation
DE pending
PCT pending
Reference ID
14/058TLB
Service
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