caa a22 4500 44529194X CHVBK 20180317142530.0 cr unu---uuuuu 170323e20100401xx s 000 0 eng 10.1007/s11626-010-9290-6 doi (NATIONALLICENCE)springer-10.1007/s11626-010-9290-6 Derivation and characterisation of the human embryonic stem cell lines, NOTT1 and NOTT2 [Elektronische Daten] [Helen Priddle, Cinzia Allegrucci, Paul Burridge, Maria Munoz, Nigel Smith, Lyndsey Devlin, Cecilia Sjoblom, Sarah Chamberlain, Sue Watson, Lorraine Young, Chris Denning] The ability to maintain human embryonic stem cells (hESCs) during long-term culture and yet induce differentiation to multiple lineages potentially provides a novel approach to address various biomedical problems. Here, we describe derivation of hESC lines, NOTT1 and NOTT2, from human blastocysts graded as 3BC and 3CB, respectively. Both lines were successfully maintained as colonies by mechanical passaging on mouse embryonic feeder cells or as monolayers by trypsin-passaging in feeder-free conditions on Matrigel. Undifferentiated cells retained expression of pluripotency markers (OCT4, NANOG, SSEA-4, TRA-1-60 and TRA-1-81), a stable karyotype during long-term culture and could be transfected efficiently with plasmid DNA and short interfering RNA. Differentiation via formation of embryoid bodies resulted in expression of genes associated with early germ layers and terminal lineage specification. The electrophysiology of spontaneously beating NOTT1-derived cardiomyocytes was recorded and these cells were shown to be pharmacologically responsive. Histological examination of teratomas formed by in vivo differentiation of both lines in severe immunocompromised mice showed complex structures including cartilage or smooth muscle (mesoderm), luminal epithelium (endoderm) and neuroectoderm (ectoderm). These observations show that NOTT1 and NOTT2 display the accepted characteristics of hESC pluripotency. The Society for In Vitro Biology, 2010 Human embryonic stem cells nationallicence NOTT1 nationallicence NOTT2 nationallicence Pluripotency nationallicence Differentiation nationallicence Cardiomyocytes nationallicence Micro-electrode array nationallicence Genetic modification nationallicence Priddle Helen NURTURE, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut Allegrucci Cinzia Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut Burridge Paul Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut Munoz Maria Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut Smith Nigel Department of Cytogenetics, Centre for Medical Genetics, Nottingham City Hospital NHS Trust, NG5 1PB, Nottingham, UK aut Devlin Lyndsey NURTURE, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut Sjoblom Cecilia NURTURE, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut Chamberlain Sarah NURTURE, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut Watson Sue School of Clinical Sciences, Faculty of Medicine & Health Sciences, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut Young Lorraine Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut Denning Chris Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/3-4(2010-04-01), 367-375 1071-2690 46:3-4<367 2010 46 11626 https://doi.org/10.1007/s11626-010-9290-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-010-9290-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Priddle Helen NURTURE, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut NATIONALLICENCE 700 1- Allegrucci Cinzia Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut NATIONALLICENCE 700 1- Burridge Paul Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut NATIONALLICENCE 700 1- Munoz Maria Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut NATIONALLICENCE 700 1- Smith Nigel Department of Cytogenetics, Centre for Medical Genetics, Nottingham City Hospital NHS Trust, NG5 1PB, Nottingham, UK aut NATIONALLICENCE 700 1- Devlin Lyndsey NURTURE, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut NATIONALLICENCE 700 1- Sjoblom Cecilia NURTURE, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut NATIONALLICENCE 700 1- Chamberlain Sarah NURTURE, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut NATIONALLICENCE 700 1- Watson Sue School of Clinical Sciences, Faculty of Medicine & Health Sciences, Queen's Medical Centre, NG7 2UH, Nottingham, UK aut NATIONALLICENCE 700 1- Young Lorraine Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut NATIONALLICENCE 700 1- Denning Chris Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/3-4(2010-04-01), 367-375 1071-2690 46:3-4<367 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer