Investigations of the application of naturally occurring or induced canine stem cells for treatment of dogs
The application of regenerative medicine utilising adult or induced stem cells in a developing area of veterinary science and this project proposes the isolation or development and subsequent characterisation of canine stem cells for the treatment of disease and disorders in dogs.
Project Context:Regenerative medicine is an emerging discipline in veterinary science and it is proposed that it will be utilised as an approach for the treatment of a variety of genetic, metabolic and orthopaedic diseases and injuries. Recently, reprogramming of cells to generate induced pluripotent stem cells (iPSCs) from adult differentiated cells like skin fibroblasts has gained significant interest as it would allow autologous transplantation of pluripotent cells circumventing many of the technical and ethical issues that have been of concern in human regenerative medicine. The benefit of iPS cell approaches is that differentiated cells can be harvested in reasonably non-invasive procedures (eg. skin collection) and as a resource are not encumbered with the ethical issues of embryo creation and destruction.The nature of pluripotent cells infers that they are capable of differentiation into many other cell types and this characteristic is under investigation for treatment of a large range of diseases.Mesenchymal stem cells (MSCs) are multipotent stem cells with significant therapeutic potential in veterinary medicine. These cells reside in bone marrow, adipose tissue, blood, umbilical cord blood, amniotic fluid, teeth and foetal tissues. In normal healthy animals MSCs play a role in cellular renewal processes and can differentiate into osteocytes (bone), adipocytes (fat cells), myocytes (muscle), chondrocytes (cartilage), cardiomyocytes (heart), and neurons (nervous system). As these cells can be harvested from a number of tissues and re-administered with the ability to differentiate into a range of tissue types, autologous application of MSCs has been investigated as a cell based therapeutic agent for human and animal health. While this approach has shown some promise in initial clinical trials, the conditions required for high purity culture and unambiguous identification of these stem cells have yet to be determined. This approach has already been successfully applied to a range of diseases and disorders including cartilage and bone repair, heart regeneration following cardiac infaction, neuronal regeneration following stroke or brain trauma, and wound healing. Veterinary applications in dogs may include repair of fractures, cartilage repair, relief of osteoarthritis and treatments for hip dysplasia.
- Establish in our lab and optimize cell culture methodologies from surgical waste material (adipose tissue) based on published methods.
- Screen cell cultures for surface markers by Fluorescence Activated Cell Sorting (FACS) to optimise cell isolation procedures and population homogeneity.
- Characterise identified side populations by gene expression profiles of genes specific to MSCs lineages.
- Evaluate cell labeling techniques for subsequent in vivo application.
- Establish and optimize canine iPSCs through the traditional retrovirus mediated methodology to establish the basic morphological, immunochemical and expression characteristics of these cells in vitro.
- To investigate methodologies for formation of ‘clinic-ready’ canine iPSCs by alternate methods (non-genome integrative) and compare to retrovirus induced iPSCs.
This project is supported by the Canine Research Foundation and is open to Masters, Honours and BScVet candidates.This project does not include student scholarship support.
Techniques employed will include:Canine Primary Cell CultureCell transduction (viral or other)Cell isolation/ FACSFlow cytometryReal-time PCRFluorescence microscopyCell labeling techniques
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The opportunity ID for this research opportunity is: 1876
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