Developing scalable and standardised manufacturing methods for human pluripotent stem cells

Lead Research Organisation: University of Nottingham
Department Name: School of Medicine

Abstract

So that people can live longer and more active lives there is a need to develop new affordable and effective medicines. In some cases cells that we have within our own bodies can be used to repair damaged tissues. However, in adults, this repair mechanism is very limited and often inefficient. The cells that make up the very early developing embryo are those that can go on to make all of the cells in the human body and these so called 'stem cells' when harvested from embryos have the potential to repair many types of diseased tissue in adults. Although stem cells from embryos can now be grown in laboratories, one highly trained person can only grow a few million cells in a week. Since it takes 5 billion heart cells to repair the heart muscle of a heart-attack patient, growing these cells manually is useful for research but not for treating multiple patients in practise. This project aims to combine the expertise of both stem cell scientists and engineers, to create large-scale systems for the 'manufacture' of large numbers of stem cells so the potential of stem cell therapies can be realised. This is not a trivial task since the way we culture stem cells manually is very different from those grown in 'factories'. Therefore this project will identify the parts of the manual cell culture process that need to be improved for the large scale cost effective manufacture of stem cells for therapeutic purposes. Once enough stem cells can be routinely grown and harvested, it is more likely that they can be made into enough heart, nerve and pancreas cells to begin to treat disease.

Technical Summary

Pluripotent hESCs are a major emerging platform for a wide range of therapeutic cell based products and pharmaceutical assays, however there are major barriers to their commercial-scale production. Our multidisciplinary collaboration will improve the understanding and reliability of cell expansion for pluripotent human embryonic stem (hESC) and induced pluripotency (hIPSC) cells by: 1) investigating properties of pluripotent cells that influence their processing and scale-up using our experience of multiple cell lines and culture conditions to scope generic process conditions 2) optimising and validating automated bioprocess protocols to enable robust and reproducible manufacture of hESC-based products at commercial scales. To maximise the range of manufacturing scales that are likely to be required for e.g. pharmaceutical screening processes or regenerative medicine applications, we will develop medium scale (entirely automated 90 X T175 flask T-flask culture in the CompacT SelecT) and larger scale (for potentially up to 1000L bioreactor) systems in parallel, using the same source of highly characterised cells. The processes that we deliver will have improved cost-effectiveness over current systems and will allow standardised culture protocols to be applied to multiple human pluripotent cell lines. Statistically-designed factorial experiments, underpinned by systematic process improvement, will identify the variables in manual culture methods that affect the practicality of scaled hESC manufacture. Factorial experimentation & quality optimisation (biological function, variation & cost) of the bioprocessed cell product will be achieved through gaining an understanding of all relevant variables through a unique collaboration between stem cell biologists and bioprocess/biomanufacturing engineers.

Publications

10 25 50
 
Description Assisted with establishment of robotic platforms for stem cell culture and differentiation
Exploitation Route We continued to develop the robotic platforms ourselves, which has been successful for publication and securing further funding
Sectors Healthcare,Manufacturing, including Industrial Biotechology

 
Description These have been submitted previously to BBSRC. However, I do not have access to them as I was not PI on the grant - I am covering this return for a colleague who left the University 2 years ago
First Year Of Impact 2011
Sector Healthcare
Impact Types Economic

 
Description Asha E-term fellowship
Amount £250,000 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 12/2013 
End 11/2015
 
Description BHF Centre for Regen Med
Amount £2,500,000 (GBP)
Funding ID P47352/ 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2013 
End 09/2017
 
Description BHF MyoD grant
Amount £300,000 (GBP)
Funding ID PG/14/59/31000 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2015 
End 05/2018
 
Description BHF Programme Grant (2 pilot with Sian Harding)
Amount £475,000 (GBP)
Funding ID RG/11/19/29264 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2012 
End 04/2014
 
Description BHF programme 2014-17
Amount £1,125,000 (GBP)
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2014 
End 04/2017
 
Description EPSRC equip - seahorse
Amount £115,000 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 09/2014 
End 03/2015
 
Description EU - dave
Amount £300,000 (GBP)
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 09/2012 
End 08/2015
 
Description Heart Res UK (Divya)
Amount £147,000 (GBP)
Funding ID TRP01/12 
Organisation Heart Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2013 
End 01/2015
 
Description MRC capital equipment
Amount £714,000 (GBP)
Funding ID MR/L012618/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 10/2013 
End 03/2014
 
Description NC3Rs main panel (Viola)
Amount £515,000 (GBP)
Funding ID NC/K000225/1 
Organisation National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) 
Sector Private
Country United Kingdom
Start 02/2013 
End 02/2016
 
Description NC3Rs-CRACK-IT phase 1
Amount £100,000 (GBP)
Organisation National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) 
Sector Private
Country United Kingdom
Start 12/2013 
End 06/2014
 
Description NC3Rs-CRACK-IT phase 2
Amount £1,000,000 (GBP)
Organisation National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) 
Sector Private
Country United Kingdom
Start 01/2015 
End 01/2018
 
Description University of Nottingham Strategic Development Fund "Nottingham Regenerative Medicine Centre" 1.1.15-31.12.18. £602,140 (PI)
Amount £602,000 (GBP)
Funding ID n/a 
Organisation University of Nottingham 
Sector Academic/University
Country United Kingdom
Start 11/2014 
End 10/2020
 
Description • BHF Programme Grant RG/15/6/31436: Predicting anti-arrhythmic drug efficacy from the divergent molecular basis of RyR2 dysfunction in genetic arrhythmia syndromes
Amount £1,100,000 (GBP)
Funding ID RG/15/6/31436 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2015 
End 09/2020
 
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 • BIRAX grant 04BX14CDLG. Gene targeted optogenetics in hPSC-cardiovascular cells for transplantation into animal models of heart dysfunction. 1.9.15-31.8.18. £400K
Amount £400,000 (GBP)
Funding ID 04BX14CDLG 
Organisation British Council 
Sector Charity/Non Profit
Country United Kingdom
Start 11/2015 
End 10/2018
 
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
 
Description • EPSRC Programme Grant EP/N006615/1: 'Next Generation Biomaterials Discovery'. PI: Morgan Alexander. Multiple Co-Is. 1.10.15-30.9.20. £6.6m
Amount £6,600,000 (GBP)
Funding ID EP/N006615/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 10/2015 
End 09/2020
 
Description • MRC grant MR/M017354/1: MICA: Development of Metrics and Quality Standards for Scale up of Human Pluripotent Stem Cells. 1.5.15-30.4.17 £1.
Amount £1,200,000 (GBP)
Funding ID MR/M017354/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 05/2015 
End 04/2017
 
Description Automation (Development of robot) 
Organisation Tecan UK Ltd
Country United Kingdom 
Sector Private 
PI Contribution Bespoke robotic platform for automated culture and differentiation of human stem cells
Collaborator Contribution Role in automation
Impact Publications, grants
Start Year 2010
 
Description BIRAX interactions (Mummery, Gepstein) 
Organisation Technion - Israel Institute of Technology
Country Israel 
Sector Academic/University 
PI Contribution New partnership between three labs
Collaborator Contribution Skills in progenitor cell biology and cell transplantation in vivo
Impact Early stage at present
Start Year 2015
 
Description CRACK-IT consortium 
Organisation GlaxoSmithKline (GSK)
Department Safety Assessment GSK
Country United Kingdom 
Sector Private 
PI Contribution consortium working on safety assessment
Collaborator Contribution all active contributors to researcher and academic input
Impact publications and grants
Start Year 2014
 
Description CRACK-IT consortium 
Organisation Leiden University
Country Netherlands 
Sector Academic/University 
PI Contribution consortium working on safety assessment
Collaborator Contribution all active contributors to researcher and academic input
Impact publications and grants
Start Year 2014
 
Description CRACK-IT consortium 
Organisation Pluriomics BV
Country Netherlands 
Sector Private 
PI Contribution consortium working on safety assessment
Collaborator Contribution all active contributors to researcher and academic input
Impact publications and grants
Start Year 2014
 
Description CRACK-IT consortium 
Organisation University of Glasgow
Country United Kingdom 
Sector Academic/University 
PI Contribution consortium working on safety assessment
Collaborator Contribution all active contributors to researcher and academic input
Impact publications and grants
Start Year 2014
 
Description CRACK-IT consortium 
Organisation University of Hamburg
Country Germany 
Sector Academic/University 
PI Contribution consortium working on safety assessment
Collaborator Contribution all active contributors to researcher and academic input
Impact publications and grants
Start Year 2014
 
Description Electrophysiology mechanisms (Harmer / Tinker) 
Organisation Queen Mary University of London
Country United Kingdom 
Sector Academic/University 
PI Contribution mechanism of electrophysiology in hPSC-cardiomyocytes
Collaborator Contribution Skills in detailed electrophyioslogy
Impact grants awarded. Papers in progress
Start Year 2014
 
Description Industrial collaboration (GSK via CRACK-IT) 
Organisation GlaxoSmithKline (GSK)
Department Safety Assessment GSK
Country United Kingdom 
Sector Private 
PI Contribution Facilitated award of NC3Rs CRACK-IT grant
Collaborator Contribution Implicit in the sponsorship and construction of the challenge
Impact Financial, international collaboration, skills sharing, joint PhD students, researcher exchange
Start Year 2013
 
Description MRC-TEL consortium (Manchester, UKSCB, Sanger, TEL) 
Organisation National Institute for Biological Standards and Control (NIBSC)
Country United Kingdom 
Sector Public 
PI Contribution joint grant with industrial partner
Collaborator Contribution joint grant with industrial partner
Impact joint grant with industrial partner
Start Year 2015
 
Description MRC-TEL consortium (Manchester, UKSCB, Sanger, TEL) 
Organisation The Wellcome Trust Sanger Institute
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution joint grant with industrial partner
Collaborator Contribution joint grant with industrial partner
Impact joint grant with industrial partner
Start Year 2015
 
Description MRC-TEL consortium (Manchester, UKSCB, Sanger, TEL) 
Organisation Tokyo Electron
Country Japan 
Sector Private 
PI Contribution joint grant with industrial partner
Collaborator Contribution joint grant with industrial partner
Impact joint grant with industrial partner
Start Year 2015
 
Description MRC-TEL consortium (Manchester, UKSCB, Sanger, TEL) 
Organisation University of Manchester
Country United Kingdom 
Sector Academic/University 
PI Contribution joint grant with industrial partner
Collaborator Contribution joint grant with industrial partner
Impact joint grant with industrial partner
Start Year 2015
 
Description surface work (Morgan Alexander, Nottingham) 
Organisation University of Nottingham
Department School of Pharmacy
Country United Kingdom 
Sector Academic/University 
PI Contribution lead the stem cell work of this collaboration
Collaborator Contribution Multiple grant awards and papers
Impact grants and papers
Start Year 2010
 
Description syngenta 
Organisation Syngenta International AG
Country Global 
Sector Public 
PI Contribution stem cell automation
Collaborator Contribution intellectual and financial input
Impact advances in stem cell automation
Start Year 2009
 
Description Various presentations that Lisa, Yan and Roger did for BBSRC projects 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Presentations, workshops, posters, industrial engagement
Year(s) Of Engagement Activity 2012