title: Autophagic response to cell culture stress in pluripotent stem cells.
abstract: Autophagy is an important conserved cellular process, both constitutively as a recycling pathway for long lived proteins and as an upregulated stress response. Recent findings suggest a fundamental role for autophagic processes in the maintenance of pluripotent stem cell function. In human embryonic stem cells (hESCS), autophagy was investigated by transfection of LC3-GFP to visualize autophagosomes and with an antibody to LC3B protein. The presence of the primary cilium (PC) in hESCs as the site of recruitment of autophagy-related proteins was also assessed. HESCs (mShef11) in vitro displayed basal autophagy which was upregulated in response to deprivation of culture medium replacement. Significantly higher levels of autophagy were exhibited on spontaneous differentiation of hESCs in vitro. The PC was confirmed to be present in hESCs and therefore may serve to coordinate autophagy function.
authors.affiliation: Centre for Stem Cell Biology, Department of Biomedical Sciences, University of Sheffield, S10 2TN, United Kingdom.
authors.forename: Siân
authors.initials: S
authors.lastname: Gregory
authors.affiliation: Centre for Stem Cell Biology, Department of Biomedical Sciences, University of Sheffield, S10 2TN, United Kingdom.
authors.forename: Sushma
authors.initials: S
authors.lastname: Swamy
authors.affiliation: Centre for Stem Cell Biology, Department of Biomedical Sciences, University of Sheffield, S10 2TN, United Kingdom.
authors.forename: Zoe
authors.initials: Z
authors.lastname: Hewitt
authors.affiliation: Centre for Stem Cell Biology, Department of Biomedical Sciences, University of Sheffield, S10 2TN, United Kingdom.
authors.forename: Andrew
authors.initials: A
authors.lastname: Wood
authors.affiliation: Centre for Stem Cell Biology, Department of Biomedical Sciences, University of Sheffield, S10 2TN, United Kingdom.
authors.forename: Richard
authors.initials: R
authors.lastname: Weightman
authors.affiliation: Centre for Stem Cell Biology, Department of Biomedical Sciences, University of Sheffield, S10 2TN, United Kingdom. Electronic address: h.d.moore@shef.ac.uk.
authors.forename: Harry
authors.initials: H
authors.lastname: Moore
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D001343:Autophagy; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002923:Cilia / Q000378:metabolism; D003470:Culture Media; D053595:Embryonic Stem Cells / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D010588:Phagosomes / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology*; D014162:Transfection
chemical_list: D000906:Antibodies; D003470:Culture Media; C502070:MAP1LC3B protein, human; D008869:Microtubule-Associated Proteins; D049452:Green Fluorescent Proteins
keywords: Autophagy; LC3-GFP; Pluripotent stem; Primary cilium