αVβ5 and CD44 Are Oxygen-Regulated Human Embryonic Stem Cell Attachment Factors
Human embryonic stem cells (hESCs) have great potential for clinical therapeutic use. However, relatively little is known of the mechanisms which dictate their specificity of adhesion to substrates through adhesion proteins including integrins. Previous observations demonstrated enhanced clonogenici...
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Online Access: | http://dx.doi.org/10.1155/2013/729281 |
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doaj-d30dd56b9b74437189c35b694b676ab72020-11-24T23:02:54ZengHindawi LimitedBioMed Research International2314-61332314-61412013-01-01201310.1155/2013/729281729281αVβ5 and CD44 Are Oxygen-Regulated Human Embryonic Stem Cell Attachment FactorsDeepak Kumar0Saniya Gupta1Ying Yang2Nicholas R. Forsyth3Guy Hilton Research Centre, Institute of Science and Technology in Medicine, University of Keele, Thornburrow Drive, Hartshill, Stoke-on-Trent, Staffordshire ST4 7QB, UKGuy Hilton Research Centre, Institute of Science and Technology in Medicine, University of Keele, Thornburrow Drive, Hartshill, Stoke-on-Trent, Staffordshire ST4 7QB, UKGuy Hilton Research Centre, Institute of Science and Technology in Medicine, University of Keele, Thornburrow Drive, Hartshill, Stoke-on-Trent, Staffordshire ST4 7QB, UKGuy Hilton Research Centre, Institute of Science and Technology in Medicine, University of Keele, Thornburrow Drive, Hartshill, Stoke-on-Trent, Staffordshire ST4 7QB, UKHuman embryonic stem cells (hESCs) have great potential for clinical therapeutic use. However, relatively little is known of the mechanisms which dictate their specificity of adhesion to substrates through adhesion proteins including integrins. Previous observations demonstrated enhanced clonogenicity in reduced oxygen culture systems. Here, we demonstrated via antibody blocking experiments that αVβ5 and α6 significantly promoted hESC attachment in 2% O2 only, whereas blockage of CD44 inhibited cell attachment in 21% O2 alone. Immunofluorescence confirmed expression of αVβ5 and CD44 in both 2% O2 and 21% O2 cultured hESCs while flow cytometry revealed significantly higher αVβ5 expression in 2% O2 versus 21% O2 cultured hESCs and higher CD44 expression in 21% O2 versus 2% O2 cultured hESCs. Adhered hESCs following blockage of αVβ5 in 2% O2 displayed a reduction in nuclear colocalisation of Oct-4 and Nanog with little effect observed in 21% O2. Blockage of CD44 had the converse effect with dramatic reductions in nuclear colocalisation of Oct-4 and Nanog in 21% O2 cultured hESC which retained adherence, but not in 2% O2 cultured cells. Identification of oxygen-dependent substrate attachment mechanisms in hESCs has the potential to play a role in the development of novel substrates to improve hESC attachment and culture.http://dx.doi.org/10.1155/2013/729281 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Deepak Kumar Saniya Gupta Ying Yang Nicholas R. Forsyth |
spellingShingle |
Deepak Kumar Saniya Gupta Ying Yang Nicholas R. Forsyth αVβ5 and CD44 Are Oxygen-Regulated Human Embryonic Stem Cell Attachment Factors BioMed Research International |
author_facet |
Deepak Kumar Saniya Gupta Ying Yang Nicholas R. Forsyth |
author_sort |
Deepak Kumar |
title |
αVβ5 and CD44 Are Oxygen-Regulated Human Embryonic Stem Cell Attachment Factors |
title_short |
αVβ5 and CD44 Are Oxygen-Regulated Human Embryonic Stem Cell Attachment Factors |
title_full |
αVβ5 and CD44 Are Oxygen-Regulated Human Embryonic Stem Cell Attachment Factors |
title_fullStr |
αVβ5 and CD44 Are Oxygen-Regulated Human Embryonic Stem Cell Attachment Factors |
title_full_unstemmed |
αVβ5 and CD44 Are Oxygen-Regulated Human Embryonic Stem Cell Attachment Factors |
title_sort |
αvβ5 and cd44 are oxygen-regulated human embryonic stem cell attachment factors |
publisher |
Hindawi Limited |
series |
BioMed Research International |
issn |
2314-6133 2314-6141 |
publishDate |
2013-01-01 |
description |
Human embryonic stem cells (hESCs) have great potential for clinical therapeutic use. However, relatively little is known of the mechanisms which dictate their specificity of adhesion to substrates through adhesion proteins including integrins. Previous observations demonstrated enhanced clonogenicity in reduced oxygen culture systems. Here, we demonstrated via antibody blocking experiments that αVβ5 and α6 significantly promoted hESC attachment in 2% O2 only, whereas blockage of CD44 inhibited cell attachment in 21% O2 alone. Immunofluorescence confirmed expression of αVβ5 and CD44 in both 2% O2 and 21% O2 cultured hESCs while flow cytometry revealed significantly higher αVβ5 expression in 2% O2 versus 21% O2 cultured hESCs and higher CD44 expression in 21% O2 versus 2% O2 cultured hESCs. Adhered hESCs following blockage of αVβ5 in 2% O2 displayed a reduction in nuclear colocalisation of Oct-4 and Nanog with little effect observed in 21% O2. Blockage of CD44 had the converse effect with dramatic reductions in nuclear colocalisation of Oct-4 and Nanog in 21% O2 cultured hESC which retained adherence, but not in 2% O2 cultured cells. Identification of oxygen-dependent substrate attachment mechanisms in hESCs has the potential to play a role in the development of novel substrates to improve hESC attachment and culture. |
url |
http://dx.doi.org/10.1155/2013/729281 |
work_keys_str_mv |
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