Engineered induced pluripotent stem cell lines for phenotypic selection of cardiac cell types
Induced pluripotent stem cells (iPSC) have emerged as a powerful tool, allowing us to generate and analyse human cardiomyocytes in vitro. Furthermore, iPSCs carrying specific mutations can be derived from patients, or alternatively disease-relevant mutations can be introduced by genome-editing. Both approaches provide a “disease-in-a-dish” resource, enabling insights into the underlying pathology that contributes to cardiac disease.
When performing such experiments, one significant challenge is that the iPSC differentiation results in mixed populations of cardiomyocytes, with atrial, ventricular and nodal characteristics. This project aims to address this problem by introducing genetically encoded fluorescent labels for a particular subtype of cardiomyocyte.
Once such a reporter tool has been successfully established and validated, it will be used to study the molecular consequences of disease-related mutations on cardiac genes with the aim to elucidate complex disturbances in mechano-signalling cascades.
This project will enable the student to gain insights into iPS cell culture, differentiation, CRISPR/Cas9 genome engineering. Future work may also involve establishing different reporter transgenes for other cell types. The cardiomyocytes will be matured in 2D and 3D cultures (i.e. engineered heart tissue format) and characterised for ventricular characteristics by qPCR, Western blotting, FACS and immunofluorescence.
As well as the specific training detailed above, students will have access to a wide-range of seminars and training opportunities through the many research institutes and centres based in Oxford. Students are also able to attend the Methods and Techniques course run by the MRC Weatherall Institute of Molecular Medicine. This course runs through the year, ensuring that students have the opportunity to build a broad-based understanding of differing research techniques.
Generic skills training is offered through the Medical Sciences Division's Skills Training Programme. This programme offers a comprehensive range of courses covering many important areas of researcher development: knowledge and intellectual abilities, personal effectiveness, research governance and organisation, and engagement, influence and impact. Students are actively encouraged to take advantage of the training opportunities available to them.
The department has a successful mentoring scheme, open to graduate students, which provides an additional possible channel for personal and professional development outside the regular supervisory framework. We hold an Athena SWAN Silver Award in recognition of our efforts to support the careers of female students and staff.
|1||Denning C et al. Biochim Biophys Acta. 2016;1863(7):1728-48. doi: 10.1016/j.bbamcr.2015.10.014.|
|2||Bizy A et al. Stem Cell Res. 2013;11(3):1335-47. doi: 10.1016/j.scr.2013.09.003.|
|3||Oceguera-Yanez et al. Methods 101: 43-55. doi.org/10.1016/j.ymeth.2015.12.012|
|4||Smith AS et al. Biotechnol Adv. 2017;35(1):77-94. doi: 10.1016/j.biotechadv.2016.12.002.|