A. Salerno (Liverpool, GB)University of Liverpool Biochemistry
Presenter Of 1 Presentation
18.3.1 - Development of a new technique for directing stem cells to engraft into articular cartilage lesions
Injection of undifferentiated mesenchymal stromal/stem cells (MSCs) into osteoarthritic (OA) joints can lead to a reduction in symptoms (1,2). Inducing the engraftment of MSCs into cartilage lesions may improve their efficacy after injection. We therefore propose to target MSCs through attachment to Type II collagen gelatin. This will be achieved by coating the surface of MSCs with a mutated module of the collagen binding domain of Gelatinase A, called 222, using an improved version of a method we previously developed for coating proteins onto cell surfaces (3).
Methods and Materials
We adapted the original method for binding proteins to the surface of MSCs (3) by undertaking the coating step with cells in suspension rather than when coated on tissue culture plastic. After validation of the coating methodology by confocal imaging, 222-coated MSCs were tested for their binding to denatured Type II collagen gelatin or plastic and compared with wild type (uncoated) MSCs as a control. In subsequent preliminary experiments the location of coated and wild type MSCs was tracked after injection into mouse knees, using an antibody to human Lamin A/C.
We were able successfully to coat MSCs with FITC-labelled 222 (Figure 1) and we observed greater binding of 222-coated MSCs to type II collagen gelatin than to plastic, whereas wild type (uncoated) MSCs were found to bind more to plastic than to gelatin (Figure 2). Preliminary cell tracking studies demonstrated the feasibility of tracking human MSCs after their injection into mouse knees.
We conclude that MSCs coated with 222 are better able to bind to Type II collagen gelatin and could potentially be used to target injected cells to the cartilage lesion sites.
1. A. Vegaet al., Transplantation 99, 1681-1690 (2015).
2. Y. G. Kohet al., Arthroscopy 29, 748-755 (2013).
3. J. P. Armstronget al., Nat Commun 6, 7405 (2015).