MICA: Development of Metrics and Quality Standards for Scale up of Human Pluripotent Stem Cells

Lead Research Organisation: University of Manchester
Department Name: School of Biological Sciences

Abstract

Many diseases remain untreatable in spite of advances in medicine world-wide. In recent years there has been much interest in using human pluripotent stem cells (hPSCs), unspecialised cells which can be coaxed using molecules that are found normally in the developing body to produce any cell type. Such stem cells can therefore be used to produce particular repair tissues for patients with different diseases that are so far incurable or for damaged tissues following injury. In order for this to be possible in a manner affordable to our healthcare systems, we need to generate banks of stem cells of different human background which have different immunological tissue types. These can then be used to make desired specialised cells tailored to the patient. However at the moment we do not have reliable ways to grow the cells in large numbers without damaging them, or changing their properties so that they cannot make healthy tissues. In this proposal we will develop the conditions for generating healthy stem cells in large numbers while protecting them from damaging influences. We will generate a system that will be suitable for an automated cell factory so that the cells can be monitored and maintained in pristine stress-free conditions and expanded in a stable way enabling them to in turn generate healthy tissues for the clinic. Such a system has been developed by a company, Tokyo Electron, which now has a base in the UK. We will evaluate a whole series of different properties of these hPSCs and work out which attributes of the cells predict their health and retention of the ability to make more stem cells and form specialised cells. These properies will be compared against commercial and other alternative hPSC test systems and, in the second phase, fed back to further improve culture conditions. In this subsequent second phase we will trial the improved culture conditions with continued cell monitoring for 'good' and 'bad' cell markers in the automated cell factory. The established validated properties will be made available to other researchers so that they can use them to improve their expansion of PSCs and subsequently generate different specialised cell types for different patient needs.

Technical Summary

Expansion and banking of "off the shelf" allogeneic human pluripotent stem cell (hPSC)-derived lineages poses a realistic option for tissue repair, affordable to public/private health care systems. However, the regenerative medicine community currently lacks affordable and scalable systems. This bottleneck for translational programmes requires novel solutions as a matter of urgency, with the availability of clinical grade human embryonic stem cell, and in the near future human induced pluripotent stem cell lines. The overarching goal of this proposal is the development and qualification of a novel system for the automated scale up of hPSCs invented by Tokyo Electron (EU base in Stevenage), for subsequent differentiation into therapeutically valuable lineages. The novel, entirely enclosed, automated culture system suitable for next generation stem cell therapy manufacturing will use non-invasive image evaluation and non invasive biochemical/genetic monitoring to allow evaluation of cell health and phenotype during cell expansion. Thus, only healthy cells can be selected for continued culture or entry into therapeutic differentiation protocols. In this proposal we will develop the metrics to ensure optimal conditions for stable expanded hPSC culture, including growth rates/phenotype and karyotypic/epigenetic normality. Deep phenotyping will include cell morphology via refinement of computer algorithms, genetic (e.g.CGH array) and epigenetic profiling and both discovery and targeted secretome metabolite/proteome analysis. Pluripotency will be assured via development of high throughput novel differentiation marker analysis replacing teratoma formation. We will generate new biomarkers based on culture medium analysis (secretome, biosensor data) to refine culture conditions and generate new media/substrate combinations. We will deliver online/Cloud-based metrics analysis tools available to UK/ other stem cell communities adaptable for other scale up applications.

Planned Impact

Tissue degeneration in an aging or sick population presents an ever-increasing clinical burden for patients and healthcare systems, with costs rising rapidly. The idea that stem cell, including hPSC, products might relieve this burden is tempered by the lack of scalable culture and differentiation systems that allow cell therapies to be delivered quickly at affordable cost. Through partnership with an inwardly-investing company, Tokyo Electron Ltd (TEL), our 2 phase proposal will address this need by combining deep hPSC analysis with state of the art bioinformatics and engineering approaches to develop and validate minimum sets of biomarkers that quantifiably define cell quality standards. This will be of broad use to hPSP user communities, and will also enable TEL's development of a fully enclosed Smart Cell System that can scale up and bank quality controlled hPSCs for differentiation to valuable lineages.
Researchers in the UK regenerative medicine community will benefit from Open Access to optimised protocols and reagents for culturing, monitoring and quantifying quality criteria of hPSCs. Novel resources, knowledge and tools (e.g. biosensors) will be produced from bioinformatics analyses of deep phenotyping data derived by cell characterisation, proteomics, lipidomics, metabolomics, genetic and epigenetic analyses. These will underpin and impact, multiple UK and international initiatives: £40m Regenerative Medicine & Capital Equipment Call funded 5 RCUK and 3 BHF Platform hubs; 3 MRC clinical grade hESC derivation centres; £13m MRC-Wellcome hiPSC scheme; euro29m IMI EU Bank for hiPSC (EBISC). Researchers in these initiatives and elsewhere will benefit from the new analytical kits and biosensors that spin-out from the industrial partnerships developed. Close connection (see letter of support) and researcher exchange and training within our consortium and between these initiatives will foster the next generation of interdisciplinary scientists.
Biopharma and Instry will immediately benefit via £2.9m of inward investment from TEL into its Stevenage site and £500K cash/in-kind to this project. Consortium partners have funding and/or partnerships with Intercytex, Syngenta, GSK, AstraZeneca, Roche, Clyde Biosciences & Pluriomics. As well as partnering with the academics in this consortium, TEL is linking with UK companies such as Orla Protein (Newcastle; culture substrates), Cell Guidance Systems (Cambridge; media) and Plexus Inc. (Edinburgh; robotics). These companies will benefit from our consortium, as will those we approach for the development of quality control testing kits/assays (e.g. Stem Cell Technologies, Miltenyi Biotech, LifeTech) and matrices (e.g. Nunc, Corning and Greiner). Exploitation, IP and licencing will be via Tech Transfer Offices and TEL; see Pathways to Impact and MICA for details.
The NHS will directly link to the proposal since Manchester Academic Health Science Centre (MAHSC) is one of only five Department of Health designated AHSCs in the UK, which recognises excellence across research, innovation, education, patient service and translational medicine. Brison is employed within Central Manchester NHS Foundation Trust (CMFT), a key partner of MAHSC, wherein CMFT's remit is to translate basic research into clinical practice working with professional bodies e.g. RCPath. These routes will ensure impact in the healthcare system.
The public will benefit by being the recipients of new healthcare technologies in the future. They will be well informed since the academic partners have proven track records in engagement through channels ranging from television, radio and newspapers to public engagement days, science cafes, school education events and websites (e.g. www.eurostemcell.org).
Regulators will be kept abreast of developments through existing links via e.g. HFEA Scientific Advisory Committee (Brison & Young), MHRA (Brison), and stem cell banking and regulation (Stacey).

Publications

10 25 50

publication icon
Dixon JE (2016) Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides. in Proceedings of the National Academy of Sciences of the United States of America

publication icon
Howe CL (2019) Surface plasmon resonance imaging of excitable cells. in Journal of physics D: Applied physics

 
Description HFEA Information for Quality programme
Geographic Reach National 
Policy Influence Type Participation in advisory committee
Impact acting as advisor to Human Fertilisation and Embryology Authority Information for quality programme which aims to improve regulatory oversight of the field including stem cell derivation, and improve information to patients and for delivery of clinical IVF services.
 
Description Membership of HFEA's Scientific and Clinical Advances committee
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
Impact Advising the HFEA on changes in regulation on ART practice, embryology, embryonic stem cell biology
 
Description BHF P47352/ Centre for Regenerative Medicine
Amount £2,500,000 (GBP)
Funding ID BHF P47352 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2017 
End 09/2021
 
Description FAPESP-UoB/UoN grant with Lygia Pereira, Sao Paolo
Amount £40,000 (GBP)
Organisation São Paulo Research Foundation (FAPESP) 
Sector Public
Country Brazil
Start 01/2018 
End 06/2019
 
Description Grant funding
Amount £120,000 (GBP)
Organisation Newlife 
Sector Charity/Non Profit
Country United Kingdom
Start 08/2017 
End 05/2021
 
Description MRC Confidence in Concept
Amount £58,510 (GBP)
Organisation University of Manchester 
Sector Academic/University
Country United Kingdom
Start 11/2018 
End 07/2019
 
Description RCUK/MRC (inter-University strategic grant) UKRM platform hub Analysing and Exploiting the Stem Cell Niche.
Amount £3,500,000 (GBP)
Funding ID MR/K026666/1 
Organisation UK Regenerative Medicine Platform 
Sector Academic/University
Country United Kingdom
Start 10/2013 
End 10/2017
 
Description Stoneygate Feehally research award
Amount £251,813 (GBP)
Organisation Kidney Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2017 
End 04/2020
 
Description • BHF Special Project no. SP/15/9/31605. "Coupling gene targeted reporters with fully automated compound library screening to mature hPSC-cardiomyocytes".
Amount £1,100,000 (GBP)
Funding ID SP/15/9/31605 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2016 
End 12/2019
 
Description • EPSRC Impact Accelerator Award. A Strategic Partnership with the Australian Cooperative Research Centre for Cell Therapy Manufacturing (CRC-CTM) to Commercialise Stem Cell Culture Polymers
Amount £67,000 (GBP)
Funding ID n/a 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 11/2015 
End 02/2017
 
Title MICA standard hESC cell bank (NIBSC) 
Description Central stock of well characterised H9 hESCs for use by partners in the MICA TEE project and MTA with WiCell for lab transfer completed 
Type Of Material Cell line 
Year Produced 2015 
Provided To Others? Yes  
Impact Enhanced standardisation across the investigation by the project partners through provision of cells from the central stock 
 
Description Randox 
Organisation Randox Laboratories
Country Global 
Sector Private 
PI Contribution We are in process of preparing a joint grant application for government funding
Collaborator Contribution Collaboration will be in antibodies and antibody related know how
Impact Grant application in progress
Start Year 2017
 
Description TEL 
Organisation Tokyo Electron and FBRI Kobe
Country Japan 
Sector Private 
PI Contribution Development of new iPSC culture an passaging method; Part of MICA consortium grant
Collaborator Contribution Sharing and development of new culture and passaging method for iPSC
Impact Grant applicaion awarded 2015 MRC Mica
Start Year 2012
 
Description CMFT Xmas lecture 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Xmas Royal Society-style lecture for Central Manchester NHS Foundation Trust "The Stem Cell Revolution".

School visit requests
Year(s) Of Engagement Activity 2013
 
Description Denning: German Center for Cardiovascular Research (DZHK) workshop 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at the German Center for Cardiovascular Research (DZHK) workshop
Year(s) Of Engagement Activity 2017
 
Description Denning: INCOR inter-laboratory exchange workshop 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at INCOR inter-laboratory exchange workshop, Sao Paolo, Brazil
Year(s) Of Engagement Activity 2017
 
Description Denning: University of Sao Paolo inter-laboratory exchange workshop 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at University of Sao Paolo inter-laboratory exchange workshop
Year(s) Of Engagement Activity 2017
 
Description Guardian newspaper article on ART child health 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Guardian newspaper article on ART child health
Year(s) Of Engagement Activity 2015
 
Description Heart Research UK booklet Jan 2018 - Outstanding Researcher Award 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Heart Research UK booklet Jan 2018 - Outstanding Researcher Award
Year(s) Of Engagement Activity 2017
 
Description Invited conference presentation during 2016-2017 reporting period 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presented at >20 international conferences in Europe, Middle East, Asia and USA, each having 50 to 500 delegates from professional academic, public, media, patient and industry
Year(s) Of Engagement Activity 2016,2017
 
Description Invited lecture to EU Student Parliament debate on embryo research 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact Invited lecture to EU Student parliament debate on human embryo research including embryonic stem cell derivation, in Salford, UK
Year(s) Of Engagement Activity 2016
 
Description Lancashire Humanists 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 70 people attenced a public lecture on stem cell biology

Requests for other talks to Humanist groups.
Year(s) Of Engagement Activity 2013
 
Description Membership of ESHRE ART Culture medium working party 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact ESHRE Culture medium working party established to consider safety of ART culture media, publish a report and liaise with industry over implications.
Year(s) Of Engagement Activity 2016
 
Description Membership of HFEA's Information for Quality Programme 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Policymakers/politicians
Results and Impact Membership of HFEA's Information for Quality Programme
Year(s) Of Engagement Activity 2015
 
Description Membership of HFEA's Scientific and Clinical Advances Advisory Committee 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Policymakers/politicians
Results and Impact Membership of HFEA's Scientific and Clinical Advances Advisory Committee
Year(s) Of Engagement Activity Pre-2006,2006,2007,2008,
 
Description New Scientist Article on ART child health 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Interview for article in New Scientist on research into ART child health
Year(s) Of Engagement Activity 2016
 
Description Presentation: Denning: British cardiovascular Society, Manchester, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Presentation: Denning: British cardiovascular Society, Manchester, UK
Year(s) Of Engagement Activity 2017
 
Description Radio 4 interview re 'Promising' stem cell trials for blindness 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Daniel Brison interviewed and commented on news story re stem cell therapies

Raising public awareness
Year(s) Of Engagement Activity 2012
 
Description Radio 4 interview with Mark Porter 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Sue Kimber interviewed on Radio 4 Case Notes programme re stem cell therapies

Raised public awareness, led to doubled patient and GP interest in NWESCC
Year(s) Of Engagement Activity 2012
 
Description STEM Cell Day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Talk triggered questions and interest

Students suggested they would like to apply for bio/biomed in Manchester
Year(s) Of Engagement Activity 2014
 
Description Science Media Centre press conference 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Press conference organised by SMC to discuss press release concerning submission of clinical grade embryonic stem cell lines to the UK Stem Cell Bank

Interest from media, followup stories. Followup contacts from industry resulting directly from press release.
Year(s) Of Engagement Activity 2011
 
Description Stem Cells and Regenerative medicine from chondrocytes to kidneys 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Talk on Regenerative medicine to U3A members
Year(s) Of Engagement Activity 2018
 
Description What is Regenerative Medicine? 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact A workshop to inform Undergraduates about what regenerative medicine is and how t can expand the possibiites for Medicine ( based on our successful workshop in 2017)
Year(s) Of Engagement Activity 2018
 
Description What is Regenerative Medicine? 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact Lay type presentation explaining what the opportunities are in the sector with exemplar talks form PhD students postdoctoral researchers and research fellows
Year(s) Of Engagement Activity 2017
 
Description Wolfson Foundation promotional booklet 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Produced double page article for Wolfson Impact Campaign to promote the research and facilities to which this funding organisation contributed
Year(s) Of Engagement Activity 2017
 
Description Women in Science Becoming the Best 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Dissemaination of information advice and support for women's role and leadership in scince
Year(s) Of Engagement Activity 2016