caa a22 4500 445292164 CHVBK 20180317142531.0 cr unu---uuuuu 170323e20100401xx s 000 0 eng 10.1007/s11626-010-9305-3 doi (NATIONALLICENCE)springer-10.1007/s11626-010-9305-3 The effect of human embryonic stem cells (hESCs) long-term normoxic and hypoxic cultures on the maintenance of pluripotency [Elektronische Daten] [Vladimir Zachar, Sinha Prasad, Simon Weli, Anette Gabrielsen, Karsten Petersen, Michael Petersen, Trine Fink] The maintenance of pluripotency of human embryonic stem cells (hESCs) requires a high efficiency of self-renewal. During in vitro propagation, however, spontaneous differentiation occurs frequently, and there is also a risk of chromosomal changes. In this study, we assessed the properties of hESCs after long-term culture at ambient air and 5% oxygen growth conditions. The hESC lines were grown for up to 42 and 18mo in normoxic and hypoxic conditions, respectively, and their proliferation; expression of Oct4, SSEA1, Nanog, and Notch1; karyotype; telomerase activity; and differentiation potential in vitro were evaluated. In contrast to cultures at 20% oxygen, where the central zones of the colonies underwent spontaneous differentiation, during exposure to 5% oxygen, the hESC colonies maintained a homogenous and flat morphology that was consistent with the presence of Oct4-positive undifferentiated phenotype. Irrespective of oxygen concentration, the undifferentiated cells expressed high levels of Nanog and Oct4 transcripts, normal karyotype, and high telomerase activity. When assayed for differentiation potential, they yielded derivatives of all three embryonic germ layers. Our data thus indicate that hypoxic exposure has the capacity to sustain enhanced long-term self-renewal of hESCs. The hESC lines described in the current paper can be obtained for research purposes from the Laboratory for Stem Cell Research, Aalborg University. The Society for In Vitro Biology, 2010 Human embryonic stem cells nationallicence Hypoxia nationallicence Oxygen nationallicence Self-renewal nationallicence Pluripotency nationallicence Long-term culture nationallicence Zachar Vladimir Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg, Denmark aut Prasad Sinha Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg, Denmark aut Weli Simon Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg, Denmark aut Gabrielsen Anette Ciconia, Aarhus Private Hospital, Hoejbjerg, Denmark aut Petersen Karsten Ciconia, Aarhus Private Hospital, Hoejbjerg, Denmark aut Petersen Michael Department of Clinical Genetics, The Juliane Marie Centre, Copenhagen University Hospital, Copenhagen, Denmark aut Fink Trine Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg, Denmark aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/3-4(2010-04-01), 276-283 1071-2690 46:3-4<276 2010 46 11626 https://doi.org/10.1007/s11626-010-9305-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-010-9305-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zachar Vladimir Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg, Denmark aut NATIONALLICENCE 700 1- Prasad Sinha Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg, Denmark aut NATIONALLICENCE 700 1- Weli Simon Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg, Denmark aut NATIONALLICENCE 700 1- Gabrielsen Anette Ciconia, Aarhus Private Hospital, Hoejbjerg, Denmark aut NATIONALLICENCE 700 1- Petersen Karsten Ciconia, Aarhus Private Hospital, Hoejbjerg, Denmark aut NATIONALLICENCE 700 1- Petersen Michael Department of Clinical Genetics, The Juliane Marie Centre, Copenhagen University Hospital, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Fink Trine Laboratory for Stem Cell Research, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg, Denmark aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/3-4(2010-04-01), 276-283 1071-2690 46:3-4<276 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer