Publications by authors named "Lyn Healy"

25 Publications

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Generation of an iPSC line (CRICKi001-A) from an individual with a germline SMARCA4 missense mutation and autism spectrum disorder.

Stem Cell Res 2021 Mar 20;53:102304. Epub 2021 Mar 20.

Department of Clinical Genetics, Guy's and St Thomas' Hospital, London, UK; Neural Stem Cell Biology Lab, The Francis Crick Institute, London, UK; Department of Medical and Molecular Genetics, School of Basic & Medical Biosciences, King's College London, UK; Department of Clinical Genetics, Great Ormond Street Hospital for Children NHS Foundation Trust, London, UK. Electronic address:

Germline missense mutations in the BAF swi/snf chromatin remodeling subunit SMARCA4 are associated with neurodevelopmental disorders, including Coffin Siris Syndrome (CSS). Here, we generated an induced pluripotent stem cell line from a male patient with atypical CSS features and a de novo heterozygous missense mutation in the SMARCA4 gene (c.3607C>T, p.(Arg1203Cys)). Hair root derived keratinocytes were reprogrammed using non-integrative Sendai virus vector delivery of pluripotency factors. iPSCs generated display normal morphology and molecular karyotype, express pluripotency markers and are able to differentiate into the three germ layers.
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http://dx.doi.org/10.1016/j.scr.2021.102304DOI Listing
March 2021

The International Stem Cell Banking Initiative (ISCBI).

Stem Cell Res 2021 Feb 22;53:102265. Epub 2021 Feb 22.

Francis Crick Institute, 1 Midland Road, London, UK. Electronic address:

The International Stem Cell Banking Initiative(ISCBI) was started in 2007 to bring together the leading stem cell banks distributing human pluripotent stem cell (hPSC) lines for research and development, to discuss best practice across a range of issues from donor consent to delivery of cells for use in research, diagnostics and cell-based medicines. ISCBI holds workshops around the world and on-line and regularly publishes summaries of discussions and consensus amongst experts in stem cell biology, biobanking technology, regulation and policy making. To date, experts from more than 28 countries have contributed to ISCBI activities which are frequently run in collaboration with other stem cell organisations and has co-ordinated closely with the International Stem Cell Initiative and the hPSCreg European Commission funded database of hPSC lines and clincal trials.
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http://dx.doi.org/10.1016/j.scr.2021.102265DOI Listing
February 2021

Cores laboratories: Organization for stem cell technology advancement.

Stem Cell Res 2021 Feb 22;53:102266. Epub 2021 Feb 22.

St Jude's Children's Research Hospital, Memphis, TN 38105, USA.

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http://dx.doi.org/10.1016/j.scr.2021.102266DOI Listing
February 2021

IGF1-mediated human embryonic stem cell self-renewal recapitulates the embryonic niche.

Nat Commun 2020 02 7;11(1):764. Epub 2020 Feb 7.

Human Embryo and Stem Cell Laboratory, The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.

Our understanding of the signalling pathways regulating early human development is limited, despite their fundamental biological importance. Here, we mine transcriptomics datasets to investigate signalling in the human embryo and identify expression for the insulin and insulin growth factor 1 (IGF1) receptors, along with IGF1 ligand. Consequently, we generate a minimal chemically-defined culture medium in which IGF1 together with Activin maintain self-renewal in the absence of fibroblast growth factor (FGF) signalling. Under these conditions, we derive several pluripotent stem cell lines that express pluripotency-associated genes, retain high viability and a normal karyotype, and can be genetically modified or differentiated into multiple cell lineages. We also identify active phosphoinositide 3-kinase (PI3K)/AKT/mTOR signalling in early human embryos, and in both primed and naïve pluripotent culture conditions. This demonstrates that signalling insights from human blastocysts can be used to define culture conditions that more closely recapitulate the embryonic niche.
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http://dx.doi.org/10.1038/s41467-020-14629-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7005693PMC
February 2020

Cell Lines as Biological Models: Practical Steps for More Reliable Research.

Chem Res Toxicol 2019 09 15;32(9):1733-1736. Epub 2019 Jun 15.

American Type Culture Collection (ATCC) , Manassas , Virginia 20110 , United States.

Research in toxicology relies on models such as cell lines. These living models are prone to change and may be described in publications with insufficient information or quality control testing. This article sets out recommendations to improve the reliability of cell-based research.
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http://dx.doi.org/10.1021/acs.chemrestox.9b00215DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6748863PMC
September 2019

Science-based assessment of source materials for cell-based medicines: report of a stakeholders workshop.

Regen Med 2018 12 29;13(8):935-944. Epub 2018 Nov 29.

Centre for Biological Engineering, Loughborough University, Loughborough, LE11 3TU, UK.

Human pluripotent stem cells (hPSCs) have the potential to transform medicine. However, hurdles remain to ensure safety for such cellular products. Science-based understanding of the requirements for source materials is required as are appropriate materials. Leaders in hPSC biology, clinical translation, biomanufacturing and regulatory issues were brought together to define requirements for source materials for the production of hPSC-derived therapies and to identify other key issues for the safety of cell therapy products. While the focus of this meeting was on hPSC-derived cell therapies, many of the issues are generic to all cell-based medicines. The intent of this report is to summarize the key issues discussed and record the consensus reached on each of these by the expert delegates.
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http://dx.doi.org/10.2217/rme-2018-0120DOI Listing
December 2018

Authentication of M14 melanoma cell line proves misidentification of MDA-MB-435 breast cancer cell line.

Int J Cancer 2018 02 10;142(3):561-572. Epub 2017 Oct 10.

International Cell Line Authentication Committee (ICLAC).

A variety of analytical approaches have indicated that melanoma cell line UCLA-SO-M14 (M14) and breast carcinoma cell line MDA-MB-435 originate from a common donor. This indicates that at some point in the past, one of these cell lines became misidentified, meaning that it ceased to correspond to the reported donor and instead became falsely identified (through cross-contamination or other means) as a cell line from a different donor. Initial studies concluded that MDA-MB-435 was the misidentified cell line and M14 was the authentic cell line, although contradictory evidence has been published, resulting in further confusion. To address this question, we obtained early samples of the melanoma cell line (M14), a lymphoblastoid cell line from the same donor (ML14), and donor serum preserved at the originator's institution. M14 samples were cryopreserved in December 1975, before MDA-MB-435 cells were established in culture. Through a series of molecular characterizations, including short tandem repeat (STR) profiling and cytogenetic analysis, we demonstrated that later samples of M14 and MDA-MB-435 correspond to samples of M14 frozen in 1975, to the lymphoblastoid cell line ML14, and to the melanoma donor's STR profile, sex and blood type. This work demonstrates conclusively that M14 is the authentic cell line and MDA-MB-435 is misidentified. With clear provenance information and authentication testing of early samples, it is possible to resolve debates regarding the origins of problematic cell lines that are widely used in cancer research.
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http://dx.doi.org/10.1002/ijc.31067DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5762610PMC
February 2018

Acquisition and Reception of Primary Tissues, Cells, or Other Biological Specimens.

Authors:
Lyn E Healy

Methods Mol Biol 2017 ;1590:17-27

The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.

The use and banking of biological material for research or clinical application is a well-established practice. The material can be of human or non-human origin. The processes involved in this type of activity, from the sourcing to receipt of materials, require adherence to a set of best practice principles that assure the ethical and legal procurement, traceability, and quality of materials.
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http://dx.doi.org/10.1007/978-1-4939-6921-0_3DOI Listing
February 2018

Rapid establishment of the European Bank for induced Pluripotent Stem Cells (EBiSC) - the Hot Start experience.

Stem Cell Res 2017 04 7;20:105-114. Epub 2017 Mar 7.

Pfizer Ltd (Neusentis), The Portway Building, Granta Park, Great Abington, Cambridge, CB21 6GS, UK.

A fast track "Hot Start" process was implemented to launch the European Bank for Induced Pluripotent Stem Cells (EBiSC) to provide early release of a range of established control and disease linked human induced pluripotent stem cell (hiPSC) lines. Established practice amongst consortium members was surveyed to arrive at harmonised and publically accessible Standard Operations Procedures (SOPs) for tissue procurement, bio-sample tracking, iPSC expansion, cryopreservation, qualification and distribution to the research community. These were implemented to create a quality managed foundational collection of lines and associated data made available for distribution. Here we report on the successful outcome of this experience and work flow for banking and facilitating access to an otherwise disparate European resource, with lessons to benefit the international research community. ETOC: The report focuses on the EBiSC experience of rapidly establishing an operational capacity to procure, bank and distribute a foundational collection of established hiPSC lines. It validates the feasibility and defines the challenges of harnessing and integrating the capability and productivity of centres across Europe using commonly available resources currently in the field.
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http://dx.doi.org/10.1016/j.scr.2017.03.002DOI Listing
April 2017

Ensuring the Quality of Stem Cell-Derived In Vitro Models for Toxicity Testing.

Adv Exp Med Biol 2016;856:259-297

SEURAT-1 Stem Cell Group, Paris, France.

Quality control of cell cultures used in new in vitro toxicology assays is crucial to the provision of reliable, reproducible and accurate toxicity data on new drugs or constituents of new consumer products. This chapter explores the key scientific and ethical criteria that must be addressed at the earliest stages of developing toxicology assays based on human pluripotent stem cell (hPSC) lines. It also identifies key considerations for such assays to be acceptable for regulatory, laboratory safety and commercial purposes. Also addressed is the development of hPSC-based assays for the tissue and cell types of greatest interest in drug toxicology. The chapter draws on a range of expert opinion within the European Commission/Cosmetics Europe-funded alternative testing cluster SEURAT-1 and consensus from international groups delivering this guidance such as the International Stem Cell Banking Initiative. Accordingly, the chapter summarizes the most up-date best practices in the use and quality control of human Pluripotent Stem Cell lines in the development of in vitro toxicity assays from leading experts in the field.
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http://dx.doi.org/10.1007/978-3-319-33826-2_11DOI Listing
June 2017

Reconsidering pluripotency tests: do we still need teratoma assays?

Stem Cell Res 2013 Jul 26;11(1):552-62. Epub 2013 Mar 26.

SET Foundation, Frankfurt am Main, Germany.

The induction of teratoma in mice by the transplantation of stem cells into extra-uterine sites has been used as a read-out for cellular pluripotency since the initial description of this phenomenon in 1954. Since then, the teratoma assay has remained the assay of choice to demonstrate pluripotency, gaining prominence during the recent hype surrounding human stem cell research. However, the scientific significance of the teratoma assay has been debated due to the fact that transplanted cells are exposed to a non-physiological environment. Since many mice are used for a result that is heavily questioned, it is time to reconsider the teratoma assay from an ethical point of view. Candidate alternatives to the teratoma assay comprise the directed differentiation of pluripotent stem cells into organotypic cells, differentiation of cells in embryoid bodies, the analysis of pluripotency-associated biomarkers with high correlation to the teratoma forming potential of stem cells, predictive epigenetic footprints, or a combination of these technologies. Each of these assays is capable of addressing one or more aspects of pluripotency, however it is essential that these assays are validated to provide an accepted robust, reproducible alternative. In particular, the rapidly expanding number of human induced pluripotent stem cell lines, requires the development of simple, affordable standardized in vitro and in silico assays to reduce the number of animal experiments performed.
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http://dx.doi.org/10.1016/j.scr.2013.03.001DOI Listing
July 2013

The ToxBank Data Warehouse: Supporting the Replacement of In Vivo Repeated Dose Systemic Toxicity Testing.

Mol Inform 2013 Jan 17;32(1):47-63. Epub 2013 Jan 17.

Douglas Connect, Zeiningen, Switzerland.

The aim of the SEURAT-1 (Safety Evaluation Ultimately Replacing Animal Testing-1) research cluster, comprised of seven EU FP7 Health projects co-financed by Cosmetics Europe, is to generate a proof-of-concept to show how the latest technologies, systems toxicology and toxicogenomics can be combined to deliver a test replacement for repeated dose systemic toxicity testing on animals. The SEURAT-1 strategy is to adopt a mode-of-action framework to describe repeated dose toxicity, combining in vitro and in silico methods to derive predictions of in vivo toxicity responses. ToxBank is the cross-cluster infrastructure project whose activities include the development of a data warehouse to provide a web-accessible shared repository of research data and protocols, a physical compounds repository, reference or "gold compounds" for use across the cluster (available via wiki.toxbank.net), and a reference resource for biomaterials. Core technologies used in the data warehouse include the ISA-Tab universal data exchange format, REpresentational State Transfer (REST) web services, the W3C Resource Description Framework (RDF) and the OpenTox standards. We describe the design of the data warehouse based on cluster requirements, the implementation based on open standards, and finally the underlying concepts and initial results of a data analysis utilizing public data related to the gold compounds.
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http://dx.doi.org/10.1002/minf.201200114DOI Listing
January 2013

Production and validation of a good manufacturing practice grade human fibroblast line for supporting human embryonic stem cell derivation and culture.

Stem Cell Res Ther 2012 Mar 28;3(2):12. Epub 2012 Mar 28.

NorthEast England Stem Cell Institute, Centre for Life, Times Square, Newcastle upon Tyne NE1 4EP, UK.

Introduction: The development of reproducible methods for deriving human embryonic stem cell (hESC) lines in compliance with good manufacturing practice (GMP) is essential for the development of hESC-based therapies. Although significant progress has been made toward the development of chemically defined conditions for the maintenance and differentiation of hESCs, efficient derivation of new hESCs requires the use of fibroblast feeder cells. However, GMP-grade feeder cell lines validated for hESC derivation are not readily available.

Methods: We derived a fibroblast cell line (NclFed1A) from human foreskin in compliance with GMP standards. Consent was obtained to use the cells for the production of hESCs and to generate induced pluripotent stem cells (iPSCs). We compared the line with a variety of other cell lines for its ability to support derivation and self-renewal of hESCs.

Results: NclFed1A supports efficient rates (33%) of hESC colony formation after explantation of the inner cell mass (ICM) of human blastocysts. This compared favorably with two mouse embryonic fibroblast (MEF) cell lines. NclFed1A also compared favorably with commercially available foreskin fibroblasts and MEFs in promoting proliferation and pluripotency of a number of existing and widely used hESCs. The ability of NclFed1A to maintain self-renewal remained undiminished for up to 28 population doublings from the master cell bank.

Conclusions: The human fibroblast line Ncl1Fed1A, produced in compliance with GMP standards and qualified for derivation and maintenance of hESCs, is a useful resource for the advancement of progress toward hESC-based therapies in regenerative medicine.
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http://dx.doi.org/10.1186/scrt103DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3392772PMC
March 2012

Standardization of pluripotent stem cell cultures for toxicity testing.

Expert Opin Drug Metab Toxicol 2012 Feb 17;8(2):239-57. Epub 2012 Jan 17.

Institute for Health & Consumer Protection, Systems Toxicology Unit, Joint Research Centre, European Commission, Ispra, Italy.

Introduction: Pluripotent stem cell (PSC) lines offer a unique opportunity to derive various human cell types that can be exploited for human safety assessments in vitro and as such contribute to modern mechanistically oriented toxicity testing.

Areas Covered: This article reviews the two major types of PSC cultures that are currently most promising for toxicological applications: human embryonic stem cell lines and human induced PSC lines. Through the review, the article explains how these cell types will improve the current safety evaluations of chemicals and will allow a more efficient selection of drug candidates. Additionally, the article discusses the important issues of maintaining PSCs as well as their differentiation efficiency.

Expert Opinion: The demonstration of the reliability and relevance of in vitro toxicity tests for a given purpose is mandatory for their use in regulatory toxicity testing. Given the peculiar nature of PSCs, a high level of standardization of undifferentiated cell cultures as well as of the differentiation process is required in order to ensure the establishment of robust test systems. It is, therefore, of pivotal importance to define and internationally agree on crucial parameters to judge the quality of the cellular models before enrolling them for toxicity testing.
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http://dx.doi.org/10.1517/17425255.2012.639763DOI Listing
February 2012

Screening ethnically diverse human embryonic stem cells identifies a chromosome 20 minimal amplicon conferring growth advantage.

Authors:
Katherine Amps Peter W Andrews George Anyfantis Lyle Armstrong Stuart Avery Hossein Baharvand Julie Baker Duncan Baker Maria B Munoz Stephen Beil Nissim Benvenisty Dalit Ben-Yosef Juan-Carlos Biancotti Alexis Bosman Romulo Martin Brena Daniel Brison Gunilla Caisander María V Camarasa Jieming Chen Eric Chiao Young Min Choi Andre B H Choo Daniel Collins Alan Colman Jeremy M Crook George Q Daley Anne Dalton Paul A De Sousa Chris Denning Janet Downie Petr Dvorak Karen D Montgomery Anis Feki Angela Ford Victoria Fox Ana M Fraga Tzvia Frumkin Lin Ge Paul J Gokhale Tamar Golan-Lev Hamid Gourabi Michal Gropp Guangxiu Lu Ales Hampl Katie Harron Lyn Healy Wishva Herath Frida Holm Outi Hovatta Johan Hyllner Maneesha S Inamdar Astrid Kresentia Irwanto Tetsuya Ishii Marisa Jaconi Ying Jin Susan Kimber Sergey Kiselev Barbara B Knowles Oded Kopper Valeri Kukharenko Anver Kuliev Maria A Lagarkova Peter W Laird Majlinda Lako Andrew L Laslett Neta Lavon Dong Ryul Lee Jeoung Eun Lee Chunliang Li Linda S Lim Tenneille E Ludwig Yu Ma Edna Maltby Ileana Mateizel Yoav Mayshar Maria Mileikovsky Stephen L Minger Takamichi Miyazaki Shin Yong Moon Harry Moore Christine Mummery Andras Nagy Norio Nakatsuji Kavita Narwani Steve K W Oh Sun Kyung Oh Cia Olson Timo Otonkoski Fei Pan In-Hyun Park Steve Pells Martin F Pera Lygia V Pereira Ouyang Qi Grace Selva Raj Benjamin Reubinoff Alan Robins Paul Robson Janet Rossant Ghasem H Salekdeh Thomas C Schulz Karen Sermon Jameelah Sheik Mohamed Hui Shen Eric Sherrer Kuldip Sidhu Shirani Sivarajah Heli Skottman Claudia Spits Glyn N Stacey Raimund Strehl Nick Strelchenko Hirofumi Suemori Bowen Sun Riitta Suuronen Kazutoshi Takahashi Timo Tuuri Parvathy Venu Yuri Verlinsky Dorien Ward-van Oostwaard Daniel J Weisenberger Yue Wu Shinya Yamanaka Lorraine Young Qi Zhou

Nat Biotechnol 2011 Nov 27;29(12):1132-44. Epub 2011 Nov 27.

Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, Sheffield, UK.

The International Stem Cell Initiative analyzed 125 human embryonic stem (ES) cell lines and 11 induced pluripotent stem (iPS) cell lines, from 38 laboratories worldwide, for genetic changes occurring during culture. Most lines were analyzed at an early and late passage. Single-nucleotide polymorphism (SNP) analysis revealed that they included representatives of most major ethnic groups. Most lines remained karyotypically normal, but there was a progressive tendency to acquire changes on prolonged culture, commonly affecting chromosomes 1, 12, 17 and 20. DNA methylation patterns changed haphazardly with no link to time in culture. Structural variants, determined from the SNP arrays, also appeared sporadically. No common variants related to culture were observed on chromosomes 1, 12 and 17, but a minimal amplicon in chromosome 20q11.21, including three genes expressed in human ES cells, ID1, BCL2L1 and HM13, occurred in >20% of the lines. Of these genes, BCL2L1 is a strong candidate for driving culture adaptation of ES cells.
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http://dx.doi.org/10.1038/nbt.2051DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3454460PMC
November 2011

Global solutions to the challenges of setting up and managing a stem cell laboratory.

Stem Cell Rev Rep 2012 Sep;8(3):830-43

Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur P.O, Bangalore 560064, India.

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http://dx.doi.org/10.1007/s12015-011-9326-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3412080PMC
September 2012

Stem cell banks: preserving cell lines, maintaining genetic integrity, and advancing research.

Methods Mol Biol 2011 ;767:15-27

UK Stem Cell Bank, Cell Biology and Imaging, National Institute for Biological Standards and Control, South Mimms, Hertfordshire, UK.

The ability to cryopreserve and successfully recover cell lines has been critical to the conservation of all cell lines, especially the preservation of pristine early-stage cultures and the preparation of well-characterized cell banks. Indeed, the systematic storage and establishment of cryopreserved banks of cells for the stem cell research community is fundamental to the promotion of standardisation in stem cell research and their use in clinical applications. In spite of the significant potential for the use of stem cells in research and therapy, they are challenging to maintain and have been shown to be unstable after prolonged culture that often results in permanent alterations in their genetic make-up, which ultimately alters the phenotype of the culture. This chapter will review the principles of cell bank production, techniques for the scale-up of human pluripotent stem cells, quality control, and characterisation methods for banked cell lines.
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http://dx.doi.org/10.1007/978-1-61779-201-4_2DOI Listing
November 2011

Comparison of defined culture systems for feeder cell free propagation of human embryonic stem cells.

In Vitro Cell Dev Biol Anim 2010 Apr 26;46(3-4):247-58. Epub 2010 Feb 26.

The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.

There are many reports of defined culture systems for the propagation of human embryonic stem cells in the absence of feeder cell support, but no previous study has undertaken a multi-laboratory comparison of these diverse methodologies. In this study, five separate laboratories, each with experience in human embryonic stem cell culture, used a panel of ten embryonic stem cell lines (including WA09 as an index cell line common to all laboratories) to assess eight cell culture methods, with propagation in the presence of Knockout Serum Replacer, FGF-2, and mouse embryonic fibroblast feeder cell layers serving as a positive control. The cultures were assessed for up to ten passages for attachment, death, and differentiated morphology by phase contrast microscopy, for growth by serial cell counts, and for maintenance of stem cell surface marker expression by flow cytometry. Of the eight culture systems, only the control and those based on two commercial media, mTeSR1 and STEMPRO, supported maintenance of most cell lines for ten passages. Cultures grown in the remaining media failed before this point due to lack of attachment, cell death, or overt cell differentiation. Possible explanations for relative success of the commercial formulations in this study, and the lack of success with other formulations from academic groups compared to previously published results, include: the complex combination of growth factors present in the commercial preparations; improved development, manufacture, and quality control in the commercial products; differences in epigenetic adaptation to culture in vitro between different ES cell lines grown in different laboratories.
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http://dx.doi.org/10.1007/s11626-010-9297-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2855804PMC
April 2010

International banking: checks, deposits, and withdrawals.

Cell Stem Cell 2008 Apr;2(4):305-6

In the 10 years that the technology to produce human embryonic stem cell lines has been available, hundreds of lines have been derived in numerous global locations. These cell lines are being used by researchers across diverse scientific fields to investigate the basic biology, clinical potential, and pharmaceutical applications of these cells and their progeny. In this fast-moving and rapidly growing field, how can we ensure that data generated by different laboratories using the same cell lines are comparable, reproducible, and consistent? One suggestion would be to ensure the quality of the "seed stock" material received and used by researchers. Because a number of laboratories worldwide provide stem cell lines to the scientific community, it seems logical to explore the harmonization of practices between distributors to establish cohesive standards and aid the global movement of stem cell lines to the research community. In the future, when these cells arrive in the clinic for therapeutic use, this consensus of "best practice" should ensure the consistency and facilitate the dissemination of these valuable materials.
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http://dx.doi.org/10.1016/j.stem.2008.03.007DOI Listing
April 2008

Characterization of human embryonic stem cell lines by the International Stem Cell Initiative.

Nat Biotechnol 2007 Jul 17;25(7):803-16. Epub 2007 Jun 17.

UK Stem Cell Bank, Division of Cell Biology and Imaging, National Institute for Biological Standards and Control, South Mimms, Herts., EN6 3QG, UK.

The International Stem Cell Initiative characterized 59 human embryonic stem cell lines from 17 laboratories worldwide. Despite diverse genotypes and different techniques used for derivation and maintenance, all lines exhibited similar expression patterns for several markers of human embryonic stem cells. They expressed the glycolipid antigens SSEA3 and SSEA4, the keratan sulfate antigens TRA-1-60, TRA-1-81, GCTM2 and GCT343, and the protein antigens CD9, Thy1 (also known as CD90), tissue-nonspecific alkaline phosphatase and class 1 HLA, as well as the strongly developmentally regulated genes NANOG, POU5F1 (formerly known as OCT4), TDGF1, DNMT3B, GABRB3 and GDF3. Nevertheless, the lines were not identical: differences in expression of several lineage markers were evident, and several imprinted genes showed generally similar allele-specific expression patterns, but some gene-dependent variation was observed. Also, some female lines expressed readily detectable levels of XIST whereas others did not. No significant contamination of the lines with mycoplasma, bacteria or cytopathic viruses was detected.
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http://dx.doi.org/10.1038/nbt1318DOI Listing
July 2007

The development of 'feeder' cells for the preparation of clinical grade hES cell lines: challenges and solutions.

J Biotechnol 2006 Oct 11;125(4):583-8. Epub 2006 May 11.

Division of Cell Biology and Imaging and UK Stem Cell Bank, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire EN6 3QG, United Kingdom.

The development of human embryonic stem cell (hESC) lines for research and therapy is hampered by the need to improve the basic methodologies for cell culture expansion. In most current methods hESC lines are cultured on a mouse or human feeder cell layer which appears to be the most reliable way to maintain cells stably in the undifferentiated state. However, co-culture introduces complications for studying stem cell biology and the delivery of safe therapies for the future. This article reviews the specific risks associated with any proposed clinical use of feeder cells of mouse origin and compares these with the benefits and risks of using human feeder cells. The further work required to establish clinical grade feeder cell lines for hESC line culture is significant and costly. Much work is being done to find feeder-free culture systems but these are at an early stage of development and there may be consequences that affect the value of the hESCs for research and development. These challenges should be viewed in the context of the huge amount of work that will be required over many years to develop robust differentiation protocols and establish fully defined procedures and adequate safety data for embryonic stem cell products.
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http://dx.doi.org/10.1016/j.jbiotec.2006.03.011DOI Listing
October 2006

The UK Stem Cell Bank: its role as a public research resource centre providing access to well-characterised seed stocks of human stem cell lines.

Adv Drug Deliv Rev 2005 Dec 10;57(13):1981-8. Epub 2005 Nov 10.

The UK Stem Cell Bank, Division of Cell Biology and Imaging, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, EN6 3QG, UK.

The rapidly expanding field of stem cell research offers the potential to develop therapeutic agents to treat diseases such as Parkinson's, diabetes and heart disease. It is important that stem cell lines derived from quality-controlled and well-characterised cell banks should be made available to both the scientific and clinical communities to promote high-quality research and development. The requirement in the United Kingdom (UK) for rigorous regulation of the procurement and use of embryonic stem (ES) cell lines led the UK government to fund the establishment of a national bank for stem cell lines. The UK Stem Cell Bank (UKSCB) hosted at the National Institute for Biological Standards and Control (NIBSC) is committed to working closely with the clinical and research communities to provide qualified stocks of human stem cell lines of adult, foetal and embryonic origin for both research use and for use in emerging human therapy.
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http://dx.doi.org/10.1016/j.addr.2005.07.019DOI Listing
December 2005

The histone deacetylase 9 gene encodes multiple protein isoforms.

J Biol Chem 2003 May 17;278(18):16059-72. Epub 2003 Feb 17.

Leukemia Research Fund Centre, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK.

Histone deacetylases (HDACs) perform an important function in transcriptional regulation by modifying the core histones of the nucleosome. We have now fully characterized a new member of the Class II HDAC family, HDAC9. The enzyme contains a conserved deacetylase domain, represses reporter activity when recruited to a promoter, and utilizes histones H3 and H4 as substrates in vitro and in vivo. HDAC9 is expressed in a tissue-specific pattern that partially overlaps that of HDAC4. Within the human hematopoietic system, expression of HDAC9 is biased toward cells of monocytic and lymphoid lineages. The HDAC9 gene encodes multiple protein isoforms, some of which display distinct cellular localization patterns. For example, full-length HDAC9 is localized in the nucleus, but the isoform lacking the region encoded by exon 7 is in the cytoplasm. HDAC9 interacts and co-localizes in vivo with a number of transcriptional repressors and co-repressors, including TEL and N-CoR, whose functions have been implicated in the pathogenesis of hematological malignancies. These results suggest that HDAC9 plays a role in hematopoiesis; its deregulated expression may be associated with some human cancers.
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http://dx.doi.org/10.1074/jbc.M212935200DOI Listing
May 2003

Chromosome translocations and covert leukemic clones are generated during normal fetal development.

Proc Natl Acad Sci U S A 2002 Jun 4;99(12):8242-7. Epub 2002 Jun 4.

Leukaemia Research Fund Centre for Cell and Molecular Biology, Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, UK.

Studies on monozygotic twins with concordant leukemia and retrospective scrutiny of neonatal blood spots of patients with leukemia indicate that chromosomal translocations characteristic of pediatric leukemia often arise prenatally, probably as initiating events. The modest concordance rate for leukemia in identical twins ( approximately 5%), protracted latency, and transgenic modeling all suggest that additional postnatal exposure and/or genetic events are required for clinically overt leukemia development. This notion leads to the prediction that chromosome translocations, functional fusion genes, and preleukemic clones should be present in the blood of healthy newborns at a rate that is significantly greater than the cumulative risk of the corresponding leukemia. Using parallel reverse transcriptase-PCR and real-time PCR (Taqman) screening, we find that the common leukemia fusion genes, TEL-AML1 or AML1-ETO, are present in cord bloods at a frequency that is 100-fold greater than the risk of the corresponding leukemia. Single-cell analysis by cell enrichment and immunophenotype/fluorescence in situ hybridization multicolor staining confirmed the presence of translocations in restricted cell types corresponding to the B lymphoid or myeloid lineage of the leukemias that normally harbor these fusion genes. The frequency of positive cells (10(-4) to 10(-3)) indicates substantial clonal expansion of a progenitor population. These data have significant implications for the pathogenesis, natural history, and etiology of childhood leukemia.
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http://dx.doi.org/10.1073/pnas.112218799DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC123052PMC
June 2002