Developing scalable and standardised manufacturing methods for human pluripotent stem cells
Lead Research Organisation:
University of Nottingham
Department Name: Div of Obs & Gynaecology
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.
Organisations
- University of Nottingham (Lead Research Organisation)
- Engineering and Physical Sciences Research Council (Co-funder)
- Technion - Israel Institute of Technology (Collaboration)
- Leiden University (Collaboration)
- University of Manchester (Collaboration)
- UNIVERSITY OF NOTTINGHAM (Collaboration)
- National Institute for Biological Standards and Control (NIBSC) (Collaboration)
- The Wellcome Trust Sanger Institute (Collaboration)
- QUEEN MARY UNIVERSITY OF LONDON (Collaboration)
- Tecan UK Ltd (Collaboration)
- Tokyo Electron (Collaboration)
- Syngenta International AG (Collaboration)
- UNIVERSITY OF GLASGOW (Collaboration)
- Universität Hamburg (Collaboration)
- Pluriomics BV (Collaboration)
- GlaxoSmithKline (GSK) (Collaboration)
Publications
Celiz AD
(2015)
Discovery of a Novel Polymer for Human Pluripotent Stem Cell Expansion and Multilineage Differentiation.
in Advanced materials (Deerfield Beach, Fla.)
Denning C
(2016)
Cardiomyocytes from human pluripotent stem cells: From laboratory curiosity to industrial biomedical platform.
in Biochimica et biophysica acta
Dick E
(2013)
Exon skipping and gene transfer restore dystrophin expression in hiPSC-cardiomyocytes harbouring DMD mutations.
in Stem cells and development
Hammad M
(2016)
Identification of polymer surface adsorbed proteins implicated in pluripotent human embryonic stem cell expansion.
in Biomaterials science
Matsa E
(2012)
In vitro uses of human pluripotent stem cell-derived cardiomyocytes.
in Journal of cardiovascular translational research
Matsa E
(2014)
Allele-specific RNA interference rescues the long-QT syndrome phenotype in human-induced pluripotency stem cell cardiomyocytes.
in European heart journal
Rajamohan D
(2013)
Current status of drug screening and disease modelling in human pluripotent stem cells.
in BioEssays : news and reviews in molecular, cellular and developmental biology
Smith JG
(2015)
Scaling human pluripotent stem cell expansion and differentiation: are cell factories becoming a reality?
in Regenerative medicine
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 | Public |
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 | 09/2013 |
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 | 04/2015 |
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 | 03/2012 |
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 | 03/2014 |
End | 04/2017 |
Description | EPSRC equip - seahorse |
Amount | £115,000 (GBP) |
Organisation | Engineering and Physical Sciences Research Council (EPSRC) |
Sector | Public |
Country | United Kingdom |
Start | 08/2014 |
End | 03/2015 |
Description | EU - dave |
Amount | £300,000 (GBP) |
Organisation | European Commission |
Sector | Public |
Country | European Union (EU) |
Start | 08/2012 |
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 | Public |
Country | United Kingdom |
Start | 09/2013 |
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 | Public |
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 | Public |
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 | Public |
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 | 09/2015 |
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 | Public |
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 | Public |
Country | United Kingdom |
Start | 09/2015 |
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 | Public |
Country | United Kingdom |
Start | 04/2015 |
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 |
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 | Switzerland |
Sector | Private |
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 |