For Research Use Only. Not for Use in Diagnostic Procedures.
Collaborative Phenotyping: HipSci and the Stem Cell Hotel, Davide Danovi, Centre for Stem Cells and Regenerative Medicine, King’s College London
Human pluripotent stem cells provide a virtually unlimited reservoir of homogeneous cells and are highly potent, offering considerable potential in gene discovery, disease modelling and drug development. The human induced pluripotent stem cell initiative (HipSci) brings together partners in genomics, proteomics, cell biology and clinical genetics to create a national induced pluripotent stem cells (iPSC) resource.
A robust high-content assay using the PerkinElmer Operetta instrument has been developed by the HipSci cell phenotyping group for the analysis of human iPSC lines to capture and quantify phenotypic features of iPSCs exposed to different extracellular matrix conditions. This approach is being developed to characterise iPSCs from multiple donors, and create novel platforms that can be tailored for disease modelling and drug discovery.
In this assay, cells are dissociated and seeded as single cells onto 96-well plates coated with fibronectin at different concentrations. After live imaging, cells are fixed and stained, followed by image acquisition and end-point analysis. This method allows assessment of cell number, proliferation, morphology (i.e. area, roundness and width to length of both cells and nuclei) and context feature capturing intercellular adhesion properties, and can be used to define phenotypes with live imaging and end-point based high content imaging.
Phenotypic features are found to vary between single cells and cells in contact with others. Fibronectin conditions mediate distinct responses affecting several phenotypic features (e.g. cell and nuclear morphology).
The HipSci initiative is attempting to benchmark donor variance in relation to healthy stem cells. Preliminary analysis and integration with genomics suggests that genetics are responsible for a portion of the variance observed between donors, and these corresponding genes can then be isolated.
Reference: Leha, Moens, Meleckyte, Culley, Gervasio, Kerz, Reimer, Cain, Streeter, Folarin, Stegle, Kielty, Durbin, Watt, Danovi. A high-content platform to characterise human induced pluripotent stem cell lines. Methods. 2016 Mar 1;96:85-96. doi: 10.1016/j.ymeth.2015.11.012. Epub 2015 Nov 25.