title: Derivation of vascular endothelial cells from human embryonic stem cells under GMP-compliant conditions: towards clinical studies in ischaemic disease.
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.
authors.affiliation: 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, UK.
authors.forename: A
authors.initials: A
authors.lastname: Kaupisch
authors.forename: L
authors.initials: L
authors.lastname: Kennedy
authors.forename: V
authors.initials: V
authors.lastname: Stelmanis
authors.forename: B
authors.initials: B
authors.lastname: Tye
authors.forename: N M
authors.initials: NM
authors.lastname: Kane
authors.forename: J C
authors.initials: JC
authors.lastname: Mountford
authors.forename: A
authors.initials: A
authors.lastname: Courtney
authors.forename: A H
authors.initials: AH
authors.lastname: Baker
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D001709:Biotechnology / Q000592:standards*; D015820:Cadherins / Q000378:metabolism; D018929:Cell Culture Techniques / Q000592:standards*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation; D042783:Endothelial Cells / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation; D005434:Flow Cytometry / Q000592:standards; D018507:Gene Expression Regulation, Developmental; D019983:Guideline Adherence / Q000592:standards*; D017408:Guidelines as Topic / Q000592:standards*; D006801:Humans; D007511:Ischemia / Q000473:pathology / Q000503:physiopathology / Q000601:surgery; D018919:Neovascularization, Physiologic; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D013997:Time Factors; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; D015820:Cadherins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D000099152:Cadherin 5