High throughput screening of predicted cell potency in cell-based regenerative medicine

Background: At the Institute for Science & Technology in Medicine (ISTM) Keele University/ RJAH Orthopaedic Hospital - Oswestry, cell therapy has been performed for over 10yrs with the treatment of cartilage injury by autologous chondrocyte implantation. The procedure uses cells that have been isolated from small cartilage biopsies and then increased in number via monolayer culture. The outcome is that 81% of patients show an increase in function within ~1yr, a benefit which is retained for at least 5 yrs with on-going follow-up. Another problematic orthopaedic condition, the non-healing of fractured bone (so called fracture non union), has been associated with the local presence of bone marrow cells which have a lower than normal osteoblastic function. Replacement of these aberrant cells with autologous mesenchymal stem cells (MSC) has provided a second line of cell therapy at the hospital, with good clinical outcome. Following this success, new cell therapies are being considered for patients with osteoarthritis, spinal cord injury (SCI) or pressure ulcers. Several key questions, also important to the established procedures, need to be answered prior to advancement of these new therapies. These include determining which cells are safe? Which are most effective? And, just as importantly, which can be delivered to the patient in a clinically relevant timescale and modality? The industrial partnership: We recently entered into collaboration with Imagen-Biotech Ltd (a UK SME) in a BBSRC funded short-term Industrial Interchange (2009). Our aim was to explore the use of 'high content' biology that Imagen-Biotech technology provides to assay various aspects of MSC potency. MSC are of considerable scientific interest as they are a poorly characterised heterogeneous cell populations, with intra- and inter-patient variability reported in terms of their function, which nonetheless have application in each of the future cell therapies being considered. The results of this Interchange have been very promising, e.g. we have shown that the high content methods provided a time efficient means of screening the neurotrophic activity of different MSC populations. Such efficiency is essential to the optimisation of MSC cell therapies for some neurological treatments, e.g. following SCI. Aims of the project: The purpose of this BBSRC-CASE studentship is to investigate more fully the use of high content/ high throughput screening in new and emerging cell therapies in regenerative medicine. We will examine a variety of cell types relevant to those therapies being developed in ISTM Keele/ Oswestry, including human MSC, chondrocytes, fibroblasts, keratinocytes. The studentship will provide a platform for basic research into cell biology, imaging techniques and the computational analysis/ modelling required for the screening mechanism. Phase and fluorescence microscopy will be used to quantitate the following generic characteristics for heterogeneous cell populations: 1.clonality (e.g. CFU-F) 2.cell proliferation rate (e.g. Ki-67, BRDU incorp) 3.cell senescence (e.g. SA beta gal activity, telomere length) 4.cell death/ apoptosis (e.g. TUNEL/ caspase) 5.karyotype stability 6.spontaneous cell transformation (e.g. p21/ p53/ telomerase expression) Additional analysis will examine particular traits re: cell function, e.g. variation in differententiation of MSC sub-populations; the stem cell characteristic of culture-expanded chondrocytes; the response of keratinocytes or neural cells to MSC secreted cytokines. The student will develop research skills in a unique biomedical setting, where it is possible to test whether cell characteristics determined via high content screening relate to functional outcome following cell transplantation, e.g. whether the proliferative potential or differentiation capacity of MSC to form osteoblasts in vitro correlates with their capacity to stimulate bone healing in fracture non union patients.