Strategy for the creation of clinical grade hESC line banks that HLA-match a target population

Authors

  • Laureen Jacquet,

    1. Embryonic Stem Cell Laboratories, Guy's Assisted Conception Unit, Division of Women's Health, King's College School of Medicine, London, UK
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    • These authors contributed equally to this work.

  • Emma Stephenson,

    1. Embryonic Stem Cell Laboratories, Guy's Assisted Conception Unit, Division of Women's Health, King's College School of Medicine, London, UK
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    • These authors contributed equally to this work.

  • Robert Collins,

    1. Clinical Transplantation Laboratory, GSTS Pathology, Guy's Hospital, London, UK
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    • These authors contributed equally to this work.

  • Heema Patel,

    1. Guy's and St. Thomas' Centre for Preimplantation Genetic Diagnosis and Genetics Centre, Guy's and St. Thomas' NHS Foundation Trust, London, UK
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  • Jane Trussler,

    1. Guy's and St. Thomas' Centre for Preimplantation Genetic Diagnosis and Genetics Centre, Guy's and St. Thomas' NHS Foundation Trust, London, UK
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  • Roaa Al-Bedaery,

    1. Embryonic Stem Cell Laboratories, Guy's Assisted Conception Unit, Division of Women's Health, King's College School of Medicine, London, UK
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  • Pamela Renwick,

    1. Guy's and St. Thomas' Centre for Preimplantation Genetic Diagnosis and Genetics Centre, Guy's and St. Thomas' NHS Foundation Trust, London, UK
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  • Caroline Ogilvie,

    1. Guy's and St. Thomas' Centre for Preimplantation Genetic Diagnosis and Genetics Centre, Guy's and St. Thomas' NHS Foundation Trust, London, UK
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  • Robert Vaughan,

    1. Clinical Transplantation Laboratory, GSTS Pathology, Guy's Hospital, London, UK
    2. MRC Centre for Transplantation, King's College London, King's Heath Partners, London, UK
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  • Dusko Ilic

    Corresponding author
    1. Embryonic Stem Cell Laboratories, Guy's Assisted Conception Unit, Division of Women's Health, King's College School of Medicine, London, UK
    • Tel: +44 20 7188 0547; Fax: +44 20 7188 0490

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Abstract

Here, we describe a pre-derivation embryo haplotyping strategy that we developed in order to maximize the efficiency and minimize the costs of establishing banks of clinical grade hESC lines in which human leukocyte antigen (HLA) haplotypes match a significant proportion of the population. Using whole genome amplification followed by medium resolution HLA typing using PCR amplification with sequence-specific primers (PCR-SSP), we have typed the parents, embryos and hESC lines from three families as well as our eight clinical grade hESC lines and shown that this technical approach is rapid, reliable and accurate. By employing this pre-derivation strategy where, based on HLA match, embryos are selected for a GMP route on day 3–4 of development, we would have drastically reduced our cGMP laboratory running costs.

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