Mesenchymal stem cell extraction from human umbilical cord tissue: processing to understand and minimise variability in cell yield
thesisposted on 22.11.2013, 15:26 by Andreea D. Iftimia-Mander
Human tissue banks are a potential source of cellular material for the emerging cellbased therapy industry; umbilical cord tissue (UCT) private banking is increasing in such facilities as a source of mesenchymal stem cells for future therapeutic use. However, early handling of UCT is relatively uncontrolled due to the clinical demands of the birth environment and subsequent transport logistics. It is therefore necessary to develop extraction methods that are robust to real world operating conditions,rather than idealised operation. This will be critical for all processes using primary tissue or cell sources. The research work undertaken in this PhD project was initiated by the collaboration with one of the leading private cord blood banks in the UK and later driven by the prospect of expanding the cell therapy and business potential of the bank. The investigation described in this thesis has focused on: - Developing an extraction method for human mesencymal stem cells (hMSCs) from UCT. - Understanding and minimizing the noticed variability in cell yield extracted from UCT by mapping the operating environment and assessing the risk factors before empirically determining their effect on the process. - Establishing the necessary process controls in the production of high quality hMSCs, through a series of wet experiments, targeted at narrowing down the sources of variability down to sub-process level. - Finding a novel method for assessing the cell content and viability of cords prior to processing. Therefore, helping the tissue processing facility to predict the risk of suboptimal cell yield from a given cord tissue section and processing method, given different operating ranges. - Determining the tissue storage requirements and isolation method with acceptable risk of adequate cell recovery. - Characterization of cells extracted from UCT via different extraction methods and comparison to primary cells extracted from other tissue sources. - Investigation of cryopreservation method for UCT. The result of this work provides a solid example of the type of data and analysis that will be required to inform a Quality-by-Design type approach for cell product development and manufacture. It will help tissue processing facilities and banks to predict the probability of cell yields from tissue sections given different operating ranges, and to aid and inform the experimental approach of others.
Future Health Technologies; EPSRC
- Mechanical, Electrical and Manufacturing Engineering