Characterisation of a novel pluripotent stem cell survival compound

Human pluripotent stem cells (hPSC) such as human embryonic stem cells (hESC) and induced pluripotent stem cells (hiPSC) are incredibly valuable tools for investigations within a number of scientific fields including developmental biology, toxicology, pharmacology and perhaps most importantly, regenerative medicine. HPSC have an unlimited capacity for self-renewal which allows the expansion of clinically relevant cell numbers from a relatively small supply of starting material. Furthermore, hPSC are pluripotent, meaning they retain the capacity to differentiate into all the somatic cell types within the human body. In order for the huge potential of hPSC to be realised, many hurdles must first be overcome. The most basic of these is the development of consistent and scalable culture systems that allow sufficient expansion of hPSC without the loss of the stem cell identity. Critical to this matter is the susceptibility of hPSC to apoptosis upon enzymatic disaggregation wherein approximately 80% of hPSC begin the process of apoptosis. Recent efforts to overcome this issue have focussed on the Rho associated coiled-coil kinase (ROCK) inhibitor Y27632. However there is increasing evidence that the use of Y27632 can lead to an increased risk of karyotypic instability, a decrease in proliferative capacity and a reduced capacity to differentiate in to specific cell types such as haematopoietic cell types. The work presented within this thesis describes the characterisation of T16, a novel hPSC survival compound which does not inhibit ROCK and has novel mechanism of action