Standardisation and other projects
Our group at the UK Stem Cell Bank (UKSCB) is involved in several standardisation projects.
International Stem Cell Initiative 3 (ISCI-3)
This is a continuation of previous projects funded by the International Stem Cell Forum.
The project’s aim is to test the hypothesis that an in vitro, controlled assay is predictive of an in vivo teratoma assay result, currently used as the ‘gold standard’, for the assessment of pluripotency of human pluripotent stem cells (hPSCs). If so, it could be used as a surrogate assay for assessing pluripotency in large numbers of stem cell isolates and in routine experimental research work to reduce the use of animals and enable routine confirmation of the potential pluripotency of cells, to give greater confidence in stem cell research data.
This will be assessed by a comparison of:
- xenograft teratoma assays
- embryoid body (EB) differentiation in vitro using defined growth conditions
- short-term differentiation assays using colony analysis on a specified micro-patterned surface
- Pluritest – an in silico assay – to test the ability of the gene expression pattern of the undifferentiated cells to predict differentiation potential as judged by teratomas and Ebs
Once these protocols have been shown to be fit for purpose they will be incorporated into the routine workflow of the UKSCB as part of the characterisation of the PSC lines.
Results of the project should be available in late 2014.
The UKSCB has taken part in two previous phases of the ISCI work (ISCI-1 and ISCI-2), which have carried out standardised research in the characterisation, culture and genetic stability of hPSCs. UKSCB is one of 4 international hub labs testing the protocols for robustness, reliability and reproducibility before the project is expanded to other international laboratories.
Toxicology projects
Embryonic stem-cell-based novel alternative testing strategies (ESNATS)
ESNATS is an EU Framework 7 project aimed at developing a novel toxicity test platform based on embryonic stem cells – particularly human stem cells (hESCs) with the aims of:
- accelerating drug development
- reducing related research and development costs
- proposing a powerful alternative to animal tests
We have worked as a partner tasked with providing methods for automating scale-up of stem cell production and cell banking and the qualification of assay procedures.
In the final stages of this project, the Bank took the lead on the standardisation of protocols selected by the ESNATS steering committee.
We used a practical assessment to address the effects of inter-laboratory transfer on the robustness, reproducibility and reliability of two cytotoxicity assay systems.
The ESNATS final brochure presents the main achievements and results of the ESNATS project which was concluded in October 2013.
Stem cells for relevant efficient extended and normalized toxicology (Scr&Tox)
Scr&Tox is another EU Framework 7 project. It is aimed at taking advantage of the key properties of stem cells – their ability to expand indefinitely and from there to differentiate potentially into any cell type in the human body.
These properties enable the development of a wide variety of human-cell-based test systems to use in safety evaluation in the SEURAT-1 cluster of projects.
SEURAT – Safety Evaluation Ultimately Replacing Animal Testing – is aimed at developing a long-term strategy for research and development which will lead to risk assessments based on robust pathway analysis rather than using animals to assess repeated dose toxicity of chemicals.
The UKSCB leads the biological resources work package in the Scr&Tox project. This package provides a central source of well-characterised ‘seed stock’ cells for use by the SEURAT-1 partners, enabling them to work with the same cells on a number of projects across the cluster.
To date the UKSCB has prepared banks of well-characterised hESC for this project and has recently produced a number of induced pluripotent stem cell (iPSC) lines using mRNA reprogramming technologies, which does not integrate into the genome.
These lines are available for our partners in the SEURAT-1 cluster to provide a standard line for assay development.
We are also actively involved in the SEURAT-1 Standardisation Group, setting standards for the development of cell-based assays funded by Scr&Tox and another SEURAT-1 cluster project – ToxBank.
ToxBank
ToxBank is also an EU Framework 7 project belonging to the SEURAT-1 project cluster.
It provides:
- a dedicated data warehouse for toxicity data management and modelling
- a ‘gold standards’ compound database and repository of selected test compounds
- a reference resource for cells, cell lines and tissues of relevance for in vitro systemic toxicity research
ToxBank supports the data management and analytical activities carried out across the Alternative Testing Strategies SEURAT-1 programme.
The workflow of the project is being developed to manage and provide access to all in vitro and in silico protocols and experimental data across SEURAT-1 to support an integrated data analysis.
An additional key component is the biomaterial wiki which contains data on suppliers of biomaterials so that scientists can select high quality materials for their research, which can be translated for industry safety assessment purposes.
The role of the UKSCB in this project is to lead the cell and tissue bank work package, developing a virtual biobank with links to key resources. This has involved establishing:
- the materials requirements for all the partners in the multiple projects within the SEURAT-1 cluster
- criteria and procedures for delivery of high-quality and acceptable sources of materials for toxicology testing and control
- a registry of key suppliers of hPSCs which provides information on key quality criteria and ethical provenance
So far, the project has delivered:
- best practice documents for working with PSC
- an evaluation process for suppliers of hPSCs
- an inventory and map of European suppliers for materials, resources, facilities and standards
The UKSCB is moving forward to populate the database with protocols and reagents to facilitate the transfer of assay systems for use in the SEURAT-1 cluster and is working with other partners to speed up the development of a comprehensive data warehouse.
The European Bank for induced pluripotent stem cells
The EBiSC project is an Innovative Medicines Initiative Joint Undertaking. Its resources are made up of financial contribution from the EU Framework 7 programme and the European Federation of Pharmaceutical Industries and Associations (EFPIA) companies’ ‘in kind’ contribution.
It was designed to address the increasing demand by induced Pluripotent Stem Cell (iPSC) researchers for quality-controlled, disease-relevant, research-grade iPSC lines, data and cell services.
Its goal is to demonstrate an operational banking and distribution service of iPSC lines after 3 years and after this to establish a centralised, not-for-profit bank for Europe providing all qualified users with access to scalable, cost-efficient and customised products.
Our role is to lead the training work package. This will provide a programme of formal theoretical and practical training that will contribute to standardisation of iPSC-related methodologies and promote quality standards within and outside the consortium.
Disseminating EBiSC standards and skills involves establishing a QC and characterisation regime that meets fundamental requirements for the scientific quality of iPSC lines under appropriate and proportionate levels of quality assurance to relevant industry standards. This will help deliver high quality, cost-effective and rapid scientific characterisation of banked iPSC lines.
As well as taking a lead role in the two work packages, UKSCB will also play a key advisory role on:
- governance of bio-banking best practice
- governance and regulation of human tissue
- cryopreservation
- coordination of banking procedural issues
- selection, evaluation and qualification of analytical methods for stem cell cultures
The European Human Pluripotent Stem Cell Registry (hESCreg)
The European Human Pluripotent Stem Cell Registry (hESCreg) project was originally established under EU Framework-6 to provide a central European registry of all human embryonic stem cell lines, their characteristics and evidence of their ethical provenance.
Current funding under EU framework-7 will extend the registry beyond embryonic stem cell lines to all pluripotent stem cells whatever their cellular source.
The aim is to promote access to ethically-sourced hPSC lines and to provide transparency about their characteristics and provenance.
The new project which started in June 2013 is designed to provide clear and transparent criteria for registration, qualification and validation of hPSC, and tools for communication and dissemination of information on the cell lines in the registry.
A dynamic registry of validated and qualified hPSC lines will provide baseline data to improve the efficiency of research, increase reproducibility and avoid repetition of research and unnecessary derivation of new lines.
The database will promote harmonisation, accuracy and validation of cell-related information and provide comprehensive and standardised information about the registered hPSC lines.
Strict quality requirements, based on identified and internationally accepted scientific standards, will improve transparency, standardisation and acceptance of the PSC field for stakeholders and the general public.
Our role is to develop a robust set of standard operating procedures under a code of practice for the operation of the database.
The project has a high-level advisory board of national representatives from around the world to help assure the veracity of data held and provide valuable information on national initiatives, regulations and law.
hESCreg is a freely accessible global registry for hPSC-lines. It allows searching for cell lines and information available about them. New cell lines can be registered and information can be added to cell lines that are already registered. This database is also coordinated with the EBiSC and ToxBank projects.