title: A thermoresponsive and chemically defined hydrogel for long-term culture of human embryonic stem cells.
abstract: Cultures of human embryonic stem cell typically rely on protein matrices or feeder cells to support attachment and growth, while mechanical, enzymatic or chemical cell dissociation methods are used for cellular passaging. However, these methods are ill defined, thus introducing variability into the system, and may damage cells. They also exert selective pressures favouring cell aneuploidy and loss of differentiation potential. Here we report the identification of a family of chemically defined thermoresponsive synthetic hydrogels based on 2-(diethylamino)ethyl acrylate, which support long-term human embryonic stem cell growth and pluripotency over a period of 2-6 months. The hydrogels permitted gentle, reagent-free cell passaging by virtue of transient modulation of the ambient temperature from 37 to 15 °C for 30 min. These chemically defined alternatives to currently used, undefined biological substrates represent a flexible and scalable approach for improving the definition, efficacy and safety of human embryonic stem cell culture systems for research, industrial and clinical applications.
authors.affiliation: EaStChem, School of Chemistry, University of Edinburgh, Joseph Black Building, West Mains Road, Edinburgh EH9 3JJ, UK.
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.forename: Heidi K
authors.initials: HK
authors.lastname: Mjoseng
authors.forename: Marieke A
authors.initials: MA
authors.lastname: Hoeve
authors.forename: Nina G
authors.initials: NG
authors.lastname: Bauer
authors.forename: Steve
authors.initials: S
authors.lastname: Pells
authors.forename: Rut
authors.initials: R
authors.lastname: Besseling
authors.forename: Srinivas
authors.initials: S
authors.lastname: Velugotla
authors.forename: Guilhem
authors.initials: G
authors.lastname: Tourniaire
authors.forename: Ria E B
authors.initials: RE
authors.lastname: Kishen
authors.forename: Yanina
authors.initials: Y
authors.lastname: Tsenkina
authors.forename: Chris
authors.initials: C
authors.lastname: Armit
authors.forename: Cairnan R E
authors.initials: CR
authors.lastname: Duffy
authors.forename: Martina
authors.initials: M
authors.lastname: Helfen
authors.forename: Frank
authors.initials: F
authors.lastname: Edenhofer
authors.forename: Paul A
authors.initials: PA
authors.lastname: de Sousa
authors.forename: Mark
authors.initials: M
authors.lastname: Bradley
mesh_terms: D055592:Biophysical Phenomena / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D003114:Colony-Forming Units Assay; D003470:Culture Media / Q000494:pharmacology; D004338:Drug Combinations; D004591:Electrophoresis, Polyacrylamide Gel; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry* / Q000494:pharmacology*; D007797:Laminin / Q000494:pharmacology; D011509:Proteoglycans / Q000494:pharmacology; D013314:Stress, Mechanical; D013696:Temperature; D013997:Time Factors
chemical_list: D003470:Culture Media; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D003094:Collagen