Primary cell isolation and culture, including endothelial stem cells and bone marrow. Tissue culture of multiple different cell lines. Development of cell based assays for cell viability, reactive oxygen species assays, metabolic function assays, and environmental stress (including metal ions, oxidative stress and inflammation). Use of scratch, migration and Matrigel assays for assessment of angiogenesis and endothelial cell function. Culture of cells on novel scaffolds to assess the role of the matrix composition and stiffness on cell functions. Production and purification of extracellular vesicles (microvesicles and exosomes). Production and titration of high titre lentiviral vector by QPCR, P24 ELISA and flow cytometry. Transduction and transfection of primary cells and cell lines with plasmids, retroviral and lentiviral vector.
Transcriptomic analysis with pathway analysis. Multivariate comparison of environmental, genetic and gene expression data.
Production of recombinant protein in bacterial and human cells and purification by FPLC and HPLC. ELISA, western blot, enzyme assays, glycosaminoglycan assays (Blyscan, AMAC) and functional complement assays. ICP-MS metal ion analysis.
Plasmid and genomic DNA and RNA (mRNA, miRNA rRNA) isolation, PCR, QPCR, southern blot, gel electrophoresis, genotyping, cloning, digestion, ligation, transformation and sequencing. Production of novel expression and reporter constructs.
Bone marrow transplant including models of neurodegenerative lysosomal storage disorders, tail vein bleed, intravenous, intraperitoneal and subcutaneous injections, PET scanning. Mouse behavioral analysis to assess neurodegeneration: open field test, elevated plus maze, hanging bar and inverted screen. Breeding and colony management.
Human eyes, urine, saliva, blood and cord blood processing and record maintenance in accordance with HTA. Statistical analysis of patient data.
Frozen or fixed tissue sectioning of brain, eye and other tissues. Histological stains, antibody stains, confocal and laser capture microdissection.