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Derivation of Vascular Endothelial Cells from Human Embryonic Stem Cells Under GMP-Compliant Conditions: Towards Clinical Studies in Ischaemic Disease

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Abstract

Revascularisation of ischaemic tissue remains an area of substantial unmet clinical need in cardiovascular disease. Strategies to induce therapeutic angiogenesis are therefore attractive. Our recent focus has been on human embryonic stem cell (hESC) strategies since hESC can be maintained in a pluripotent state or differentiated into any desired cell type, including endothelial cells (EC), under defined differentiation culture conditions. We recently published a protocol for non-good manufacturing practice (GMP) feeder- and serum-free hESC-EC-directed monolayer differentiation to vascular EC demonstrating the potential to generate hESC-derived EC in a GMP-compliant manner suitable for use in clinical trials. In this study we modified that laboratory protocol to GMP compliance. EC production was confirmed by flow cytometry, qRT-PCR and production of vascular structures in Matrigel®, yielding approximately 30 % mature VE-cadherin+/PECAM-1+ cells using the GMP-compliant hESC line RC13. In conclusion, we have successfully demonstrated the production of vascular EC under GMP-compliant conditions suitable for clinical evaluation.

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Abbreviations

GMP:

Good manufacturing practice

EC:

Endothelial cells

hESC:

Human embryonic stem cells

EPC:

Endothelial progenitor cells

N-CAM:

Neuronal cell adhesion molecule

KDR:

Kinase insert domain receptor

PAD:

Peripheral arterial disease

CLI:

Critical limb ischaemia

BM:

Bone marrow

EBs:

Embryoid bodies

MEF:

Mouse embryonic fibroblasts

ATMP:

Advanced therapy medicinal products

bFGF:

Basic fibroblast growth factor

VEGF:

Vascular endothelial growth factor

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Acknowledgments

We would like to thank the Technological Strategy Board and the British Heart Foundation for supporting this work. AHB is supported by the British Heart Foundation Chair of Translational Cardiovascular Sciences.

Ethical Standards

These studies were performed with project approval from the UK Stem Cell Bank Steering Committee and in accordance with current guidelines for the use of human embryonic stem cells.

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Authors and Affiliations

  1. BHF Glasgow Cardiovascular Research Centre, Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, 126 University Place, Glasgow, G12 8TA, UK

    A. Kaupisch, N. M. Kane, J. C. Mountford & A. H. Baker

  2. Roslin Cells Ltd., Scottish Centre for Regenerative Medicine, Edinburgh bioQuarter, 5 little France Drive, Edinburgh, EH16 4UU, UK

    L. Kennedy, V. Stelmanis, B. Tye & A. Courtney

Authors
  1. A. Kaupisch
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  2. L. Kennedy
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  3. V. Stelmanis
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  6. J. C. Mountford
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  8. A. H. Baker
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Correspondence to A. H. Baker.

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Kaupisch, A., Kennedy, L., Stelmanis, V. et al. Derivation of Vascular Endothelial Cells from Human Embryonic Stem Cells Under GMP-Compliant Conditions: Towards Clinical Studies in Ischaemic Disease. J. of Cardiovasc. Trans. Res. 5, 605–617 (2012). https://doi.org/10.1007/s12265-012-9379-2

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  • DOI: https://doi.org/10.1007/s12265-012-9379-2

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