A novel characterisation and separation technique for pluripotent human embryonic and hematopoeitic stem cells

Lead Research Organisation: University of Edinburgh
Department Name: Royal (Dick) School of Veterinary Scienc

Abstract

Realising the promised benefits of stem cells and their derivatives in regenerative medical therapies or in high throughput screening platforms for drug development necessitates the development of tools to purify cells to homogeneity. Ideally, such tools should be non-invasive with a capacity for separation several orders of magnitude beyond current methods. Industrial systems for the purification of therapeutic proteins provide precedents for large scale bioprocessing. To date, the vast majority of these have focused on column chromatography to achieve desired levels of purity. However, the size, sensitivity and complexity of cells present particular challenges to the downstream engineer which are unlikely to be solved by any further evolution or modification of traditional column chromatographic techniques. Consequently, novel techniques are needed. This proposal intends to develop a novel, simple, scalable and commercially useful technique for the separation/purification of human stem cells irrespective of their tissue of origin. This will be achieved by using Atomic Force Microscopy to define the topography and electrical charge distribution on the surfaces of human embryonic and adult haematopoietic stem cells followed by the use of this information in computational models to design complimentary surfaces. Surface prototypes will then be used in small-scale experimental work with living cells to demonstrate their ability to reversibly bind and separate cells and their subsequent viability. By focusing on both charge and topography of stem cell-surfaces the project will produce an adsorption-based separation technology more specifically suited for cell purification than current chromatographic techniques. The use of both adult and embryonic stem cell populations will exemplify the utility of this technology to both current and future clinical and research applications.

Technical Summary

This proposal intends to develop a scalable separation/purification technique for human stem cells capable of non-invasive and reversible processing of cells that will supercede current methods based on flow cytometry, centrifugation or magnetic separation. By using Atomic Force Microscopy with charged tips to generate charge maps the research will identify surface electrical charge differences associated with specific human stem cell populations, namely embryonic and adult haematopoietic (CD34+) stem cells. This information will be used to computationally model complimentary interactive substrates providing reversible affinity. Small-scale experimental work will be used to evaluate substrate prototypes for their capacity to enrich for stem cell populations without altering their viability or pluripotency, assessed using a range of standardised in vitro assays for cell molecular marker expression, and differentiation potential. By focusing on both charge and topography of surfaces the work will produce an adsorption-based separation technology more specifically suited for cell purification than current chromatographic techniques. This novel technology should be scalable to processing larger quantities of cells (10 E9-10) than current methods, thus making it a commercially useful solid-phase separation technique.

Publications

10 25 50
 
Description Industrial project management and development experience in support of commercial development of technology to separate human stem and derivative cells on the basis of their elasticity.
Exploitation Route Device to positively or negatively purify stem and derivative cells being manufactured for industrial or therapeutic purposes.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description They have been used to inform the development of a device to separate/purify human stem and derivative cells.
First Year Of Impact 2014
Sector Pharmaceuticals and Medical Biotechnology
Impact Types Economic

 
Title ELASTSEP 
Description A means to scalably discriminate cells on the basis of variation in cell elasticity without recourse to cell labelling. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Label-free cell discrimination. 
 
Description EBiSC 
Organisation National Institute for Biological Standards and Control (NIBSC)
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Public 
PI Contribution Consensus standards for human pluripotent stem cell production and quality control
Collaborator Contribution Consensus standards for human pluripotent stem cell production and quality control
Impact EC/IMI funded European Bank for Induced Stem Cells. I lead workpackage establishing foundational collection whilst collaborator leads on quality control WP.
Start Year 2006
 
Description RepairHD 
Organisation Cardiff University
Department Brain repair group
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Good Manufacturing Practice Grade Human pluripotent stem cell culture and qualification.
Collaborator Contribution Neuronal differentiation and animal models with which to evaluate physiological bioactivity associated with hESC derived neuronal populations.
Impact The principle outcome constitutes an EUFP7 funded RepairHD programme which focuses on Good Manufaturing Practice grade translation and preclinical animal model verification of bioactivity of an hESC derived cell therapy product in animal models of Huntingtons Disease. My expertise in GMP, hESC biosafety and supportive technology arising from MRC, CSO and BBSRC awards, awards contributes to this collaboration/partnership.
Start Year 2013
 
Description Animal Cell Technology Industrial Platform 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Lecture to Pharmaceutical Company Scientists specializing in recombinant protein production on emerging opportunities arising from hPSC technology, generally and founded on our work.
Year(s) Of Engagement Activity 2013
 
Description BDEBATE 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Professional discussion re: power and limitations of use of human cell technologies in discovery and therapy.
Year(s) Of Engagement Activity 2015
URL http://www.bdebate.org/en
 
Description British Association for Tissue Banking 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Reported on experience in standardised production of hPSC for clinical applications.
Year(s) Of Engagement Activity 2013
 
Description DASC Stem Cell and Tissue Engineering Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Annual Danish Stem Cell Society Meeting
Year(s) Of Engagement Activity 2015
URL https://dascs.nemtilmeld.dk/5/
 
Description Edinburgh International Science Festival 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Public lecture on opportunities and limitations of current state of the art in human pluripotent stem cell technology
Year(s) Of Engagement Activity 2013
 
Description Oxford Stem Cell Symposium 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation to peers covering our contributions to hPSC field.
Year(s) Of Engagement Activity 2014
 
Description Toxbank Public Forum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Lectured on own research underpinning use of hPSC in drug toxicity screening.
Year(s) Of Engagement Activity 2015
URL http://toxbank.net/toxbank-public-forum