title: Identification of polymer surface adsorbed proteins implicated in pluripotent human embryonic stem cell expansion.
abstract: Improved biomaterials are required for application in regenerative medicine, biosensing, and as medical devices. The response of cells to the chemistry of polymers cultured in media is generally regarded as being dominated by proteins adsorbed to the surface. Here we use mass spectrometry to identify proteins adsorbed from a complex mouse embryonic fibroblast (MEF) conditioned medium found to support pluripotent human embryonic stem cell (hESC) expansion on a plasma etched tissue culture polystyrene surface. A total of 71 proteins were identified, of which 14 uniquely correlated with the surface on which pluripotent stem cell expansion was achieved. We have developed a microarray combinatorial protein spotting approach to test the potential of these 14 proteins to support expansion of a hESC cell line (HUES-7) and a human induced pluripotent stem cell line (ReBl-PAT) on a novel polymer (N-(4-Hydroxyphenyl) methacrylamide). These proteins were spotted to form a primary array yielding several protein mixture 'hits' that enhanced cell attachment to the polymer. A second array was generated to test the function of a refined set of protein mixtures. We found that a combination of heat shock protein 90 and heat shock protein-1 encourage elevated adherence of pluripotent stem cells at a level comparable to fibronectin pre-treatment.
authors.affiliation: Laboratory of Biophysics and Surface Analysis, School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD, UK. morgan.alexander.nottingham.ac.uk.
authors.forename: Moamen
authors.initials: M
authors.lastname: Hammad
authors.affiliation: Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD, UK.
authors.forename: Wei
authors.initials: W
authors.lastname: Rao
authors.affiliation: Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: James G W
authors.initials: JG
authors.lastname: Smith
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Anderson
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Langer
authors.affiliation: Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: Lorraine E
authors.initials: LE
authors.lastname: Young
authors.affiliation: Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD, UK.
authors.forename: David A
authors.initials: DA
authors.lastname: Barrett
authors.affiliation: Laboratory of Biophysics and Surface Analysis, School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD, UK. morgan.alexander.nottingham.ac.uk.
authors.forename: Martyn C
authors.initials: MC
authors.lastname: Davies
authors.affiliation: Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: Chris
authors.initials: C
authors.lastname: Denning
authors.affiliation: Laboratory of Biophysics and Surface Analysis, School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD, UK. morgan.alexander.nottingham.ac.uk.
authors.forename: Morgan R
authors.initials: MR
authors.lastname: Alexander
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation; D000066449:Human Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology; D011108:Polymers / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D011108:Polymers