MICA: Moving From Autologous to Allogeneic Cell Therapy: Developing Up-Scale GMP Protocols for Orthopaedics.

点击翻译按钮获取中文摘要

Damaged cartilage can be treated at the Robert Jones and Agnes Hunt (RJAH) Orthopaedic hospital using an approved cartilage cell (chondrocyte) therapy on the NHS called autologous chondrocyte implantation (ACI) or with 'stem cells' as part of a clinical trial. In both techniques a patient's own cells are taken from healthy cartilage or bone marrow at first surgery, grown up in the lab over 2-3 weeks and implanted back into the joint to repair the cartilage at the second surgery. This procedure is time consuming and costly; what is more, some patients' cells do not grow well or have properties which mean they can no longer heal the patient's own cartilage when implanted back. To reduce cost and improve patient outcome it would be ideal to use an 'off the shelf' treatment, whereby cells from different donors have been tested and chosen for their therapeutic potential (potency) and stored in a biobank for use in a one stage procedure. The current method of growing cells in our Good Manufacturing Practice (GMP) laboratory involves the use of several procedures which are 'open' to the air and could increase the risk of infection. In addition, traditional culture techniques utilise multiple plastic flasks for growth phases, which are labour intensive and time consuming to monitor and maintain. This project proposes the use of a state of the art semi-automated machine called a bioreactor (Quantum) in which to grow the cells. This machine has a much larger internal surface area compared to what we currently use (50-100 times larger), so allowing more cells to be grown quicker. The bioreactor units are 'closed' which means that the risk of contamination is significantly reduced. Labour and manufacturing costs per dose are also reduced considerably with the Quantum. We have carried out assessments of the Quantum bioreactor system in our laboratory for the expansion of human stem cells isolated from healthy bone marrow or umbilical cords and human chondrocytes isolated from total knee replacements. Extended characterisation of cells using specialist techniques to assess proteins and genes showed that cells grown in the Quantum and using traditional techniques displayed comparable therapeutic properties. Moreover, the Quantum generated very large stem cell numbers resulting, on average, in 132 million bone marrow stem cells being produced in 13 days and 169 million umbilical cord stem cells in 8 days, cell numbers for chondrocytes were 75 million also in 8 days (i.e. using the bioreactor produced over forty times as many stem cells and nearly twenty times as many chondrocytes as we would get with current methods).The Quantum has been used to produce stem cells in large international clinical trials treating conditions such as diabetes and kidney disease, both in the UK and elsewhere. In the current proposal we will establish GMP manufacturing protocols for cartilage cells grown in the Quantum system. The aim of the study is to grow large numbers of cells from healthy juvenile and adult tissues donated to research in order to establish the best source for treating cartilage defects and osteoarthritis. In addition, we will ensure they are at least as good as the cells we currently use by comprehensively assessing the characteristics of the cartilage and stem cells grown in the Quantum in terms of their ability to grow and produce cartilage in the laboratory and by comparing their properties to cells currently used in the clinic.