Pluripotent Stem Cell Banking
Human pluripotent stem cells (hPSCs) have the ability to self-renew indefinitely and differentiate into all cell types. This makes them an invaluable source of material for multiple applications, such as disease modeling, toxicology screening, and cell replacement therapies. These properties, along with more accessible and efficient reprogramming and genome editing techniques, have contributed to the rapid increase in the number of hPSC lines being created and used worldwide.
By 2018, it was estimated that over 10,000 hPSC lines had been reported in the literature¹ based on an analysis of more than 3,500 hPSC research papers. This figure, combined with the finding that approximately one third of the lines submitted for distribution were found to fail minimal quality criteria², raises concerns about the lack of consensus on hPSC quality attributes and reproducibility between research groups.
Newly generated or established hPSC lines must be characterized to ensure consistency and reproducibility in stem cell research across labs. Although characterization takes time and can be expensive, it is possible to spread the cost over many years and projects by creating a well-characterized cell bank. Periodically thawing and assessing new vials from the bank can boost confidence that the cells have not drifted too far from the characterized bank. As well as helping to achieve cell line standardization, a fully characterized hPSC bank ensures your supply of consistent, high-quality hPSCs as a starting point for further research.
Continue reading below for resources—including related webinars, publications, and tools—to learn how to build a characterized cell bank to ensure your supply of consistent, high-quality cells.
Key Publications
- Recurrent genetic abnormalities in hPSCs include aberrations in chromosomes 1, 12, 17, and 20. This study systematically evaluated the sensitivity of G-banding, fluorescence in situ hybridization (FISH), and quantitative and digital droplet polymerase chain reaction (PCR). The detection limits for these methods were around 5 - 20%; thus, cultures that were determined to be karyotypically normal using these methods could, in fact, harbor undetected low-level mosaicism. Baker et al. concluded, “Recognition of this point is crucial for developing strategies for routine laboratory practice as well as the regulation for the use of hPSCs in regenerative medicine.”
Baker D et al. (2016) Detecting genetic mosaicism in cultures of human pluripotent stem cells. Stem Cell Reports 7(5): 998–1012. - The International Stem Cell Initiative screened 136 hPSC lines (at early and late passage) for genetic changes occurring during routine culture. The majority (~65%) of hPSC lines retained a normal karyotype during culture, and the remaining cell lines either acquired a cytogenetic change over time or were abnormal at both early and late passage. The recurrent karyotypic abnormalities identified were similar to previous studies involving chromosomes 1, 12, 17, and 20. The authors also proposed BCL2L1, an anti-apoptotic gene, as a candidate for driving selection of variant hPSCs with a duplication of 20q11.21 in hPSCs.
International Stem Cell Initiative et al. (2011) Screening ethnically diverse human embryonic stem cells identifies a chromosome 20 minimal amplicon conferring growth advantage. Nat Biotechnol 29(12): 1132–44. - Using recommendations and guidance from the International Stem Cell Banking Initiative, this report summarizes recommendations for the minimum dataset required to consider an induced pluripotent stem cell (iPSC) line for clinical use.
Stephen S et al. (2018) Quality control guidelines for clinical-grade human induced pluripotent stem cell lines. Regenerative Med 13(7): 859–66. - The International Stem Cell Banking Initiative published this guidance document following its first meeting in October 2007. Despite being prepared from the perspective of human embryonic stem cell (hESC) culture, the information can be applied to all human stem cell lines, including iPSC lines, in many ways.
International Stem Cell Banking Initiative (2009) Consensus guidance for banking and supply of human embryonic stem cell lines for research purposes. Stem Cell Rev Rep 5(4): 301–14.
On-Demand Webinars
Improving Reproducibility of Your hPSC Research by Generating a High-Quality Cell Bank
Many hPSC lines are available, but not all of them pass quality tests. Characterizing your cells ensures that you know what you're working with and facilitates reproducibility in your research. In this seminar, Dr. Matthew Hildebrandt from STEMCELL Technologies reviews why it’s important to characterize cell lines. He also discusses how to build a well-characterized working cell bank to ensure consistent cells throughout a project and how it can help you save time and money in the long term.
- Creating a well-characterized cell bank
- Importance of characterizing hPSC lines
- Key considerations of banking, including quality attributes and hPSC culture bottlenecks
- Ensuring validity and reproducibility of data
Speaker Bio
Nature Research Round Table: Standards for Pluripotent Stem Cell Banking
Dr. Glyn Stacey, Director of the International Stem Cell Banking Initiative (ISCBI), discusses standards for PSC banking with a focus on scientific challenges for PSCs, standardization of ethics, the role of stem cell biobanks, and best practices for cell therapy. This presentation and the following Q&A session were moderated by Dr. Joanne Mountford, formerly from the University of Glasgow.
This presentation was part of a Round Table series in 2018 titled “Challenges in Ensuring hPSC Quality,” hosted by STEMCELL Technologies in partnership with Nature Research. Global experts gathered at the Springer Nature headquarters in London, UK, to tackle some of the most pertinent issues impacting the use of hPSCs, ranging from fundamental biology research to therapeutic applications. Explore the full series here.
Speaker Bio
Nature Research Round Table: Defining and Maintaining Pluripotency & hPSC Line Registration and Banking - Panel Discussion
In this panel discussion led by Dr. Ludovic Vallier, formerly from the Wellcome Trust Sanger Institute, and Dr. Joanne Mountford, formerly from the University of Glasgow, keynote speakers respond to questions from the forum and evaluate findings on topics raised in the previous talks.
This presentation was part of a Round Table series in 2018 titled “Challenges in Ensuring hPSC Quality,” hosted by STEMCELL Technologies in partnership with Nature Research. Global experts gathered at the Springer Nature headquarters in London, UK, to tackle some of the most pertinent issues impacting the use of human pluripotent stem cells (hPSCs), ranging from fundamental biology research to therapeutic applications. Explore the full series here.
Speaker Bio
More Resources
Webinar: Development, Compatibility, and Applications of mTeSR™ Plus; an Enhanced Medium for the Maintenance of Human Pluripotent Stem Cells
In this talk, Dr. Melanie Kardel is joined by Loren Ornelas and Dr. Dhruv Sareen from the Cedars-Sinai Induced Pluripotent Stem Cell (iPSC) Core. This talk features an overview of mTeSR™ Plus and core services, highlights key collaborations, and describes the core facility’s experience with mTeSR™ Plus, including cell line transition and quality control.
On-Demand Course: Expansion of hPSCs in 3D Suspension Culture
Depending on your research goals, you may need to generate large numbers of hPSCs for your cell banks. In this free, on-demand course, our in-house PSC experts demonstrate how to safely and rapidly expand and scale up hPSCs in 3D suspension culture.
Tools
Healthy Control Human iPSC Line, Female, SCTi003-A
Start your research confidently with a reliable source of healthy, high-quality induced pluripotent stem cells (iPSCs) from the SCTi003-A cell line, certified by hPSCreg® and compatible for use with TeSR™ and STEMdiff™ media.
hPSC Genetic Analysis Kit
Quickly and cost-effectively detect the 8 most common karyotypic abnormalities reported in hPSCs.
Frequently Asked Questions on iPSCs
Find answers to frequently asked questions about iPSC lines from STEMCELL.
mFreSR™
Achieve high thawing efficiencies by preserving your human ESCs and iPSCs using mFreSR™, a serum-free freezing medium compatible with mTeSR™1, TeSR™2, and mTeSR™ Plus.
CryoStor® CS10
Provide your cells and tissues with a safe, protective environment during freezing, thawing, and storage with CryoStor® CS10.
FreSR™-S
Cryopreserve your hPSCs as single cells with FreSR™-S, a defined, serum-free, and animal component-free medium compatible with mTeSR™1, TeSR™2, TeSR™-E8™, and mTeSR™ Plus.
References
- Guhr A et al. (2018) Recent Trends in Research with Human Pluripotent Stem Cells: Impact of Research and Use of Cell Lines in Experimental Research and Clinical Trials. Stem cell reports 11(2): 485–96
- Felkner D et al. (2019, June) “Human pluripotent stem cell quality: A scientific wake‐up call.” Poster presented at the ISSCR Annual Meeting, Los Angeles, California
