Embryology Unit

Lab head: Prof. Patick P.L. Tam
Location: Children's Medical Research Institute

Website: http://www.cmri.org.au/Research/Research-Units/Embryology

Deciphering mechanisms leading to self-organising differentiation of human pluripotent stem cells.

Primary supervisor: Pierre Osteil

A key requisite for the application of human induced pluripotent stem cells (hiPSCs) for disease modelling and cellular therapy is the ability to differentiate the stem cells efficiently into clinically useful cell types. In the course of optimising the protocol of directed differentiation, it becomes evident that the cell lines differ in their ability to differentiate into particular cell types. This process is driven by the chemical and physical environment.

Our study will use multiple isogenic (same patients) hiPSCs lines that will be differentiated on micropatterned dishes. This will allow us to recapitulate the embryonic environment that leads to proper gastrulation, hence the formation of the three primitive germ layers, known as the ectoderm, the mesoderm and the endoderm. The student will test several combination of growth factors and assess the efficiency of differentiation by confocal microscopy and time-lapse video microscopy. Transcriptomic analysis will be performed as well to decipher the impact of the environment on differentiation. To validate genetic interactions with lineage propensity, gene edited lines (by CRISPR/Cas9 technology) will be tested in parallel for their ability to differentiate towards the three lineages.  In summary the student will learn about maintaining the hiPSCs in culture, protein detection using immunostaining and western blot, imaging and genome editing.

This project will test the hypothesis that modulation of the activity of signalling factors and physical constraints can rewire the lineage propensity of pluripotent stem cells, with the aim to further our understanding on cell fate decision during the early development of the human embryo.

Discipline: Applied Medical Sciences, Westmead
Co-supervisors: Patrick Tam
Keywords: Stem cells, Bioinformatics, Birth defects